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Thymic stromal lymphopoietin (TSLP) is a key cytokine that initiates and promotes allergic inflammation both in humans and mice. It is well known that TSLP is important in initial step of inflammation by stimulating dendritic cells to promote Th2 differentiation of naive T cells. However, TSLP is abundantly produced in the late phase of inflammation, as well; therefore, we focused on the function of TSLP in chronic Th2-type inflammation. By establishing a novel (to our knowledge) chronic allergic skin inflammation mouse model with repetitive challenges of hapten after sensitization, we demonstrated that CD4 T cell–specific deletion of TSLP receptor (TSLPR) resulted in near-complete ablation of ear swelling and infiltration of CD4 T cells and eosinophils, but after second challenge. Of note, TSLPR deletion on CD4 T cells did not affect acute inflammation. As expected, transfer of Ag-sensitized wild-type CD4T cells, but not of TSLPR-deficient CD4T cells, increased skin inflammation in the model upon challenge. Furthermore, production of IL-4 from TSLPR-deficient CD4T cells in inflamed ear lesions was markedly diminished, demonstrating that TSLP-dependent IL-4 production from CD4T cells was critical for the exacerbation of skin inflammation. Similar results were obtained in Th2-type allergic skin inflammation model using MC903. Collectively, these results indicate that TSLP acts directly on CD4 T cells to elicit pathogenesis of Th2 cells, thereby having a critical role in exacerbation of skin inflammation in the chronic phase.

Fine regulation of the Ras/mitogen-activating protein kinase (MAPK) pathway is crucial in controlling the survival, proliferation, and development of various types of cells. Ras-activating protein-like 3 (Rasal3) is a T cell-specific Ras GTPase-activating protein that negatively regulates T cell receptor (TCR)-induced activation of Ras/MAPK pathway. Rasal3-deficient mice showed a decreased number of naive T cells because Rasal3 is required for the survival of naive T cells. In the current study, we observed ameliorated Type1 T helper (Th1) cell- and Type2 T helper (Th2) cell-dependent contact hypersensitivity reactions in Rasal3-deficient mice, along with a marked shortage of T cells at regional lymph node. Activated Rasal3-deficient T cells showed an increased cell death with reduced Bcl2 expression, suggesting that Rasal3 is required for the survival of not only naive T cells but also activated T cells. Collectively, Rasal3 controls the magnitude of inflammatory responses through the survival of both naive T cells and activated T cells in vivo.

In recent studies, better clinical outcomes for patients with left-sided colon cancer (CC) compared with right-sided CC have been reported; however, in such investigations, the chemotherapy regimens included molecular-targeting agents. To the best of our knowledge, the impact of primary tumor location as a predictive factor in patients suffering from CC treated with cytotoxic anticancer agents alone has not been investigated. The aim of the present study was to determine the impact of the primary tumor location as a predictive factor of patients undergoing the following cytotoxic anticancer agent regimens: Leucovorin and fluorouracil + oxaliplatin (FOLFOX) or Leucovorin and fluorouracil + irinotecan (FOLFIRI), using the collagen gel droplet-embedded drug sensitivity test (CD-DST). Between March 2008 and April 2017, tumor specimens were obtained from 133 patients suffering from colorectal cancer (CRC) who had not received preoperative chemotherapy. CD-DST was performed and the growth inhibition rate (IR) was determined in FOLFOX and FOLFIRI regimens. The associations between tumor location and IR values for each condition were evaluated. In the present study, the prognosis of patients receiving palliative chemotherapy as well as treatment with molecularly-targeted agents was also investigated. There were no significant differences in the IRs (%) of the two regimens using CD-DST for right-sided tumors compared with left-sided tumors, including or excluding the rectum. The median survival times of patients with right CC and left CC who had received palliative chemotherapy and treatment with molecularly-targeted agents were 960 and 1,348 days, respectively. Primary tumor location did not represent a predictive factor for the efficacy of treatment with cytotoxic anticancer agent regimens using CD-DST. However, patients suffering from left-sided CC were revealed to exhibit better clinical outcomes compared with patients suffering from right-sided CC when molecularly-targeted agent regimens were administered. Therefore, the results of the present study suggested that molecularly-targeted agents rather than cytotoxic anticancer agents may result in improved clinical outcomes for patients with CRC suffering from left-sided tumors.

ESET/SETDB1, one of the major histone methyltransferases, catalyzes histone 3 lysine 9 (H3K9) trimethylation. ESET is critical for suppressing expression of retroviral elements in embryonic stem cells; however, its role in the immune system is not known. We found that thymocyte-specific deletion of ESET caused impaired T cell development, with CD8 lineage cells being most severely affected. Increased apoptosis of CD8 single-positive cells was observed, and TCR-induced ERK activation was severely inhibited in ESET−/− thymocytes. Genome-wide comprehensive analysis of mRNA expression and H3K9 trimethylation revealed that ESET regulates expression of numerous genes in thymocytes. Among them, FcγRIIB, whose signaling can inhibit ERK activation, was strongly and ectopically expressed in ESET−/− thymocytes. Indeed, genetic depletion of FcγRIIB in ESET−/− thymocytes rescued impaired ERK activation and partially restored defective positive selection in ESET−/− mice. Therefore, impaired T cell development in ESET−/− mice is partly due to the aberrant expression of FcγRIIB. Collectively, to our knowledge, we identify ESET as the first trimethylated H3K9 histone methyltransferase playing a crucial role in T cell development.

5-Fluorouracil (5-FU) dosing has traditionally been based on the body surface area (BSA) in colorectal cancer treatment. However, there is accumulating evidence that dosing based on BSA may be of limited use. The purpose of the present study was to evaluate the changes in 5-FU plasma levels and tumor response as well as the severity of adverse events in patients with cancer treated with 5-FU combined chemotherapy. The dosing amount of 5-FU was determined based on the BSA. Blood samples were collected, and 5-FU plasma levels in 15 patients with colorectal cancer were measured three times (0, 22 and 40 h before and after the start of infusion) during constant-infusion of 5-FU for 46 h by an immunoassay. 5-FU plasma levels were significantly higher at 22 and 40 h compared with at 0 h (P

Acquired immune deficiency syndrome (AIDS) is mainly caused by infection with human immunodeficiency virus-1 (HIV-1) and still poses a global threat for which we lack a protective or therapeutic vaccine. Dendritic cells (DCs) play a major role in the onset of HIV infection, providing one of the primary sites of HIV replication, and also act as viral reservoirs in vivo. Previous studies have shown that baculovirus (BV) induces strong host immune responses against infections and malignancies. In this study, we infected human monocyte-derived DCs with recombinant BV (AcCAG-gag) and showed that AcCAG-gag-infected human DCs underwent maturation and produced interferon alpha and other proinflammatory cytokines accompanied by increases in the mRNA and protein expression levels of APOBEC3 (A3A, A3F, and A3G), proteins associated with the inhibition of HIV-1 replication. Surprisingly, HIV-1 inhibition is also observed in human DCs infected with a wild-type BV, as determined by the production of inflammatory cytokines, the expression of A3, and a reduction in the p24 level. Our findings outline the mechanism underlying the inhibition of HIV-1 in BV-infected human DCs and pave the way for the use of BV as an effective tool for immunotherapy against HIV-1.

Kimura et al. demonstrate that NQO1 plays a crucial role in degrading IκB-ζ protein through forming the complex together with PDLIM2 and selectively suppresses IL-6 and IL-12 production induced by TLR ligands., NAD(P)H:quinone oxidoreductase 1 (NQO1) protects cells against oxidative stress and toxic quinones. In this study, we found a novel role of NQO1 in suppressing Toll-like receptor (TLR)–mediated innate immune responses. NQO1-deficient macrophages selectively produced excessive amounts of IL-6, IL-12, and GM-CSF on LPS stimulation, and the deletion of NQO1 in macrophages exacerbated LPS-induced septic shock. NQO1 interacted with the nuclear IκB protein IκB-ζ, which is essential for the TLR-mediated induction of a subset of secondary response genes, including IL-6, and promoted IκB-ζ degradation in a ubiquitin-dependent manner. We demonstrated that PDLIM2, known as the ubiquitin E3 ligase, participates in NQO1-dependent IκB-ζ degradation. NQO1 augmented the association between PDLIM2 and IκB-ζ, resulting in increased IκB-ζ degradation. Collectively, this study describes a mechanism of the NQO1–PDLIM2 complex as a novel and important regulator in the innate immune signaling and suggests the therapeutic potential of NQO1 in TLR-mediated inflammation and disorders., Graphical Abstract

Irritant contact dermatitis (ICD) is associated with local release of inflammatory mediators such as reactive oxygen species and regulated by various antioxidative enzymes and antioxidants. Although Nqo1 is involved in antioxidative reactions and detoxification, its role in ICD remains unknown. Nqo1-deficient mice exhibited augmented ear swelling accompanied by neutrophil infiltration in the croton oil–induced mouse ICD model. In the skin of Nqo1-deficient mice, Vγ5Vδ1+ dendritic epidermal T cells (DETCs), which are known to suppress ICD, were severely reduced. As the transfer of DETCs into Nqo1-deficient mice reversed an increased ICD response, loss of DETCs could account for the increased ICD. DETCs from Nqo1-deficient mice were sensitive to oxidative stress–induced cell death in vitro, and antioxidant NAC treatment in the ears of these mice rescued the number of DETCs and produced a normal ICD response. Taken together, the current results demonstrate that antioxidative enzyme Nqo1 regulates ICD through DETC maintenance.

The cytokine thymic stromal lymphopoietin (TSLP) has been implicated in the initiation and progression of allergic inflammation through its ability to activate dendritic cells (DCs). However, the identity of the DC subset that responds to TSLP is not known. In this study we use a CCL17 reporter strain to identify the TSLP-responsive DC subset. In vitro, TSLP induced CD11bhigh DCs to express CCL17, to increase CCR7-mediated migration activity, and to drive Th2 differentiation of naive CD4 T cells. In vivo, following skin sensitization, we found that a subset of Ag-bearing CCL17+CD11bhigh migratory DCs, but not Ag-bearing CCL17− migratory DCs, in skin lymph nodes were capable of driving Th2 differentiation and were dramatically reduced in TSLPR-deficient mice. Taken together, these results demonstrate that TSLP activated a subset of CD11b+ DCs in the skin to produce CCL17, upregulate CCR7, and migrate to the draining lymph node to initiate Th2 differentiation.

Serum iron levels have been reported to increase following the administration of various anticancer drugs. An increase in serum iron levels during therapy with leucovorin and fluorouracil plus oxaliplatin (FOLFOX) or leucovorin and fluorouracil plus irinotecan (FOLFIRI) was also observed. The aim of this study was to investigate the correlation between serum iron levels and prognosis in advanced colorectal cancer (CRC) patients treated with FOLFOX/FOLFIRI ± molecularly-targeted drugs. Serum iron levels were measured prior to and at 48 h after treatment with FOLFOX/FOLFIRI ± molecularly-targeted drugs in 72 advanced CRC patients, all of whom succumbed to the disease between December, 2005 and February, 2012. No patients received radiotherapy. Taking the median rate of increase in serum iron levels as the cut-off value in each therapy, the patients were divided into cohort I (increase rate greater than the cut-off value in at least one therapy) or cohort II (increase rate less than the cut-off value in all therapies). The χ2 test and the t-test were used to compare patient characteristics between the two cohorts. Prognosis was evaluated between the two cohorts using the Kaplan-Meier method, the log-rank test and the Cox proportional hazards regression analysis. No significant bias in patient characteristics (including the frequency of chemotherapy or number of patients treated with molecularly-targeted drugs) was observed between the two cohorts. Serum iron levels were transiently elevated following treatment (P

Leucovorin (FOL) and fluorouracil (5-FU) plus oxaliplatin (l-OHP; FOLFOX) or FOL and 5-FU plus irinotecan (SN-38; FOLFIRI) are widely used as first-line chemotherapy regimens in the treatment of advanced colorectal cancer (CRC). However, second-line chemotherapy must be abandoned in certain cases due to disease progression, adverse effects or high medical cost. Therefore, the most effective regimen should be selected as first-line chemotherapy. We reported that individualization of first-line treatment (FOLFOX/FOLFIRI/Dual/Poor responder) was possible using the collagen gel droplet-embedded culture drug sensitivity test (CD-DST) and that individualized first-line chemotherapy with CD-DST may improve the prognosis of patients with unresectable CRC. The aim of the present prospective cohort study was to evaluate the individualization of first-line chemotherapy using CD-DST, with a focus on prognosis. Between March 2008 and December 2015, tumor specimens were obtained from 120 patients with CRC who had not received preoperative chemotherapy. CD-DST was performed and the growth inhibition rate (IR) was determined by exposure for 24 h with 5-FU and l-OHP (6.0 and 3.0 µg/ml, respectively) and 5-FU and SN-38 (6.0 and 0.2 µg/ml, respectively). The cumulative distribution of IR values under each condition was evaluated on the basis that the clinical response to FOLFOX and FOLFIRI is equivalent (~50%). The prognosis of dual responder was improved compared with that of poor responders, however this difference was identified to be significant. There was no different prognosis between patients treated with an appropriate first-line regimen and patients treated with an inappropriate first-line regimen in dual responders. However, in poor responders, there were significant differences of prognosis between patients treated with an appropriate first-line regimen and patients treated with an inappropriate first-line regimen (P=0.036). In conclusion, the results from the present study suggest that administration of the recommended first-line regimen using CD-DST for patients with unresectable CRC is important for the improvement of prognosis, particularly in poor responders.

Originally shown to promote the growth and activation of B cells, TSLP is now known to have wide-ranging impacts on hematopoietic and nonhematopoietic cell lineages, including DCs, basophils, eosinophils, mast cells, CD4+, CD8+, and NK T cells, B cells, and epithelial cells. Whereas the role of TSLP in the promotion of TH2 responses has been studied extensively in the context of lung- and skin-specific allergic disorders, it is becoming increasingly clear that TSLP may impact multiple disease states within multiple organ systems, including the blockade of TH1/TH17 responses and the promotion of cancer and autoimmunity. This review will highlight recent advances in the understanding of TSLP signal transduction, as well as the role of TSLP in allergy, autoimmunity, and cancer. Importantly, these insights into the multifaceted roles of TSLP could potentially allow for novel, therapeutic manipulations of these disorders.

The effectiveness of cetuximab (Cmab) against KRAS p.G13D mutant-type tumors has been reported. In this study, we report a case of metastatic ascending colon cancer harboring a KRAS p.G13D mutation in a 65-year-old female. Considering the absence of symptoms and the post-operative risk of respiratory system complications due to multiple lung metastases, particularly at the entrance to the left main bronchus, anticancer drug therapy was selected as first-line therapy. With informed consent, FOLFOX4 [folinic acid (FOL), fluorouracil (F) plus oxaliplatin (OX)] + Cmab therapy was administered as preoperative chemotherapy. A good preoperative response was obtained to the chemotherapy, with a metastatic lesion disappearing from the entrance to the left main bronchus. Subsequent resection was performed successfully with no post-operative complications. Although a histopathological examination of the resected tissue specimen revealed residual cancer cells, it also showed the marked efficacy of the chemotherapy regimen used. In this study, we describe a case of metastatic ascending colon cancer harboring a KRAS p.G13D mutation in which the patient responded well to first-line therapy with FOLFOX4 + Cmab.

The cytokine thymic stromal lymphopoietin (TSLP) has been implicated in the development and progression of allergic inflammation in both humans and mice. TSLP has been shown to promote a Th2-type response through upregulation of OX40L on dendritic cells, and through direct induction of IL-4 production in naive CD4+ T cells. However, its direct effect on effector Th cells has not been extensively investigated. In this study, we show that the level of TSLP receptor (TSLPR) expression on mouse effector Th2 cells is higher than on Th1 and Th17 cells, and that TSLP induced proliferation of effector Th2, but not Th1 nor Th17 cells. TSLP also induced the phosphorylation of signal transducer and activator of transcription (Stat) 5, and expression of the anti-apoptotic factor Bcl-2 in Th2 cells. Finally, TSLP-mediated proliferation on Th2 cells was enhanced by TCR stimulation, through IL-4-mediated induction of TSLPR expression. Taken together, these results indicate that TSLP is involved in exacerbation of mouse Th2-mediated allergic inflammation in a Th2 environment through direct stimulation of Th2 effector cells.

A dendritic cell (DC) vaccine strategy has been developed as a new cancer immunotherapy, but the goal of complete tumor eradication has not yet been achieved. We have previously shown that baculoviruses potently infect DCs and induce antitumor immunity against hepatomas in a mouse model. Baculovirus-infected, bone marrow-derived DCs (BMDCs) display increased surface expression of costimulatory molecules, such as CD80, CD86 and major histocompatibility complex (MHC) classes I and II, and secrete interferons and other proinflammatory cytokines. In this study, we evaluated the induction of antitumor immunity in mice by baculovirus-infected BMDCs against lung cancer and melanoma. After treatment with baculovirus-infected BMDCs, murine lung tumors caused by Lewis lung carcinoma (LLC) cells were significantly reduced in size, and the survival of the mice was improved. In addition, experiments using a melanoma mouse model showed that baculovirus-infected BMDCs inhibited tumor growth and improved survival compared with controls. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatinine levels remained normal in baculovirus-infected BMDC-treated mice. Our findings show that baculovirus-infected DCs induce antitumor immunity and pave the way for the use of this technique as an effective tool for DC immunotherapy against malignancies.

We previously reported that the baculovirus induced a strong host immune response against infections and malignancies. Among the immune cells, the dendritic cells were most strongly infected and activated by the baculovirus, although the exact mechanism remained unclear. Here, we evaluated the non-specific immune responses of bone marrow-derived dendritic cells (BMDCs) after infection by a wild-type baculovirus. MHC class I and II molecules and co-stimulation molecules (CD40, CD80, and CD86) on BMDCs were up-regulated by baculovirus infection. At the same time, the BMDCs produced pre-inflammatory cytokines (IL-6, IL12p70, and TNF-alpha) and IFN-alpha. NK cells showed IFN-gamma production, CD69 up-regulation, and enhanced cytotoxicity when they were co-cultured with baculovirus-infected BMDCs. T cells showed IFN-gamma production, CD69 up-regulation, and cell proliferation. Ex vivo analysis performed in vitro produced similar results. These findings suggested that baculovirus-infected dendritic cells induce non-specific immune responses and can be used as an immunotherapeutic agent against viral infections and malignancies, together with present therapeutic drug regimens.

FOLFOX4 and FOLFIRI are effective regimens for the treatment of advanced colorectal cancer, and their use together with molecular targeting drugs has recently become more common. In the present study, we evaluated the changes in the serum iron levels of patients undergoing FOLFOX4 or FOLFIRI therapy alone or in combination with bevacizumab (BV). The serum iron level was increased 48 h after therapy and was restored to baseline 2 weeks afterwards in colorectal cancer patients who received FOLFOX4 or FOLFIRI alone or in combination with BV. This transient increase in serum iron was observed repeatedly during chemotherapy. The serum iron level was 71.66±28.96 μg/dl (mean ± standard deviation) before treatment and significantly increased to 186.82±83.17 μg/dl (p

The purpose of this study was to determine the effect of the addition of oxaliplatin (l-OHP) or irinotecan (SN-38) to 5-fluorouracil (5-FU) using the collagen gel droplet embedded culture-drug sensitivity test (CD-DST) to establish whether leucovorin plus 5-FU should be administered in combination with l-OHP (FOLFOX) or SN-38 (FOLFIRI) in individualized first-line chemotherapy for the treatment of advanced colorectal cancer (CRC). Specimens of primary tumors were obtained from 24 CRC patients who had received no preoperative chemotherapy. CD-DST was performed, and the inhibition rate (IR) was obtained under multiple incubation conditions. The effects of addition of l-OHP or SN-38 were evaluated for the same area under the concentration curve (AUC) of 5-FU based on linear regression analysis. Approximate expression and correlation coefficients (5-FU vs. 5-FU + l-OHP, 5-FU vs. 5-FU + SN-38; AUC of 5-FU=72 and 5-FU vs. 5-FU + l-OHP, 5-FU vs. 5-FU + SN-38; AUC of 5-FU=144) were y=0.94x+8.53 (R(2)=0.95, p

Studies of human asthma and of animal models of allergic airway inflammation revealed a crucial role for Th2 cells in the pathogenesis of allergic asthma. Kruppel-type zinc finger proteins are the largest family of a regulatory transcription factor for cellular development and function. Zinc finger protein (Zfp) 35 is an 18-zinc finger motif-containing Kruppel-type zinc finger protein, while its function remains largely unknown. The aim of this study was to clarify the role of Zfp35 in the pathogenesis of Th2-dependent allergic inflammation, such as allergic asthma. We examined airway eosinophilic inflammation and hyperresponsiveness in two mouse models, which use our newly generated Zfp35-deficient (Zfp35−/−) mice and adoptive transfer of cells. In Zfp35−/− mice, Th2 cell differentiation, Th2 cytokine production, eosinophilic inflammation, and airway hyperresponsiveness were substantially enhanced. Furthermore, adoptive transfer of Ag-sensitized Zfp35−/− CD4 T cells into the asthmatic mice resulted in enhanced airway inflammation and airway hyperresponsiveness. These results indicate that Zfp35 controls Th2 cell differentiation, allergic airway inflammation, and airway hyperresponsiveness in a negative manner. Thus, Zfp35 may control Th2-dependent diseases, such as allergic asthma.

We have evaluated the 5-fluorouracil sensitivity of cancer cells from colorectal cancer patients using the collagen gel droplet embedded drug sensitivity test under multiple drug concentrations and contact durations. After converting drug concentration and contact time to the area under the curve (AUC) and plotting against the growth inhibition rate, the correlation between AUC and the growth inhibition rates was approximated to the logarithmic regression curve. In this study, to further validate the reliability of the regression curve, the growth inhibition rate was calculated from the regression curve and the actual growth inhibition rate was compared at AUC of 48 mug h/ml. No significant difference was observed in the growth inhibition rates between the two groups by paired t-test (P=0.590). A strong positive correlation was found between the two groups by regression analysis (y=0.7555x+10.514, R=0.8236). This result strongly suggests that in-vitro antitumor effect of 5-fluorouracil depends on the AUC in colorectal cancer and the AUC-inhibition rate curve is reliable. We can obtain the inhibition rate from AUC and vice versa using the AUC-inhibition rate curve. We can also calculate the individualized AUCIR50, AUC value that gives 50% growth inhibition, using the AUC-inhibition rate curve. This could be useful to establish individualized chemotherapy using the collagen gel droplet embedded drug sensitivity test.

INTRODUCTION: We previously investigated the effects of sodium ferrous citrate (SFC) on neutrophils in vitro, and revealed that SFC inhibited neutrophil functions such as superoxide production and chemotaxis. Salazosulphapyridine (SASP) and predonizolone (PSL) are widely used for treatment of ulcerative colitis (UC). In this study, we evaluated the actions of SASP or PSL on the functions of neutrophils. Moreover, we evaluated the combined effects of SASP or PSL and SFC on neutrophil functions.METHODS: Neutrophils were incubated with SASP or PSL in the absence or presence of SFC. For determination of superoxide production, neutrophils were further stimulated with phorbol myristate acetate (PMA), complement-opsonized zymosan (OPZ) or N-formyl-menthionyl-leucyl-phenylalamine (fMLP). The superoxide production was measured on the basis of superoxide dismutase-inhibitable cytochrome c. Neutrophil migration was measured by a Boyden chamber assay using zymosan-activated serum (C5a) as a chemotactic factor. To examine the expression of CD11b and CD62L, neutrophils were stimulated with fMLP, and then incubated with phycoerythrin-labeled anti-CD11b and FITC-labeled anti-CD62L monoclonal antibodies, and the expression of CD11b and CD62L were analyzed by flow cytometry.RESULTS: SASP dose-dependently inhibited the superoxide production induced by all stimuli examined, whereas, PSL did not affect the superoxide production. SASP and PSL inhibited chemotaxis toward C5a in a dose-dependent fashion. Furthermore, SASP and PSL inhibited the fMLP-induced up-regulation of CD11b and down-regulation of CD62L. Interestingly, the combinats on the SASP or PSL and SFC exerted the strong inhibitory actions on the superoxide production and neutrophil migration compared with SASP and PSL alone.CONCLUSIONS: The present observations suggest that the combined administration of SASP or PSL with SFC may have a potential to reduce mucosal injury by suppressing neutrophil functions in UC.

Background Nishiki-nezumi Cinnamon/Nagoya (NC/Nga) mice raised in nonair-controlled conventional circumstances spontaneously develop atopic dermatitis-like skin lesions; however, the underlying mechanisms remain unclear. Objective We wanted to identify the critical intracellular signaling molecules in T cells that induce atopic dermatitis-like skin legions in NC/Nga mice. Methods We examined the levels of signal transduction and cytokine production in T cells, particularly those in atopic dermatitis-like lesions induced by the topical injection of mite antigens in NC/Nga mice under specific pathogen-free conditions. Results In NC/Nga mice maintained under specific pathogen-free conditions, the capability of T H 1/T H 2 and T cytotoxic 1/T cytotoxic 2 (Tc1/Tc2) cell differentiation increased significantly. T-cell antigen receptor–mediated activation of the extracellular signal-regulated kinase/mitogen-activated protein kinase cascade and nuclear factor-κB (NF-κB) signaling were enhanced in NC/Nga T cells. The expression of T H 2 cytokines (IL-4, IL-13, and IL-5) and that of GATA-binding protein 3 (GATA3), avian musculoaponeurotic fibrosarcoma (c-Maf), NF-κB, and activator protein 1 (AP1) selectively increased in draining lymph node T cells of NC/Nga mice. Moreover, the cell transfer of inhibitory NF-κB mutant-infected T H 2 cells reduced ear thickness in the mite antigen-induced skin lesion of NC/Nga mice. Conclusion Hyperresponsive T H 2 cells with an enhanced activity of NF-κB and AP1 play a crucial role in the pathogenesis of atopic dermatitis-like skin lesions in NC/Nga mice. Clinical implications These results indicate potential therapeutic usefulness of developing selective inhibitors for NF-κB in the treatment of human atopic dermatitis.

極めて稀な大網原発多形型脂肪肉腫の1例を報告する.症例は44歳,男性,約1カ月間続く腹部膨満感を主訴に当院を受診した.初診時,心窩部に手拳大の可動性のある腫瘤を触知した.腹部超音波検査で内部不均一な8.5cmの腫瘤が描出され,腹部造影CT検査では辺縁不整で不均一に造影効果を伴う等吸収値の腫瘤が描出された.以上より腹腔内腫瘍の診断で開腹手術を施行した.主病変は大網の中央部に存在したが,播種と考えられる無数の小結節が腹腔内全体に認められたため,主病変および数個の播種結節を切除して手術を終了した.主病変は12×8×5cmで,重量約300gであった.病理組織学的に腫瘍は大網原発多形型脂肪肉腫と診断された.患者は手術から約4カ月後に腹膜播種のため死亡した.

BACKGROUND AND PURPOSE: The inflammatory bowel diseases(IBD) such as ulcerative colitis(UC) are intractable diseases of unknown origin. The mucosal injury of UC is presumed to be partly caused by reactive oxygen species(ROS) produced by neutrophils. Chronic inflammation of UC results in chronic anemia, to which iron is given. In this study, to evaluate the effect of sodium ferrous citrate(SFC) on neutrophil functions, we measured superoxide generation and migration of neutrophils treated with SFC in vitro.METHODS: Superoxide generation induced by phorbol myristate acetate(PMA), opsonized zymosan(OPZ) and N-formyl-menthionyl-leucyl-phenylalanine(fMLP) was determined based on the superoxide dismutaseinhibitable cytochrome c reduction. To further clarify the inhibitory mechanism of SFC, the membrane fraction was isolated and NADPH-dependent superoxide production was measured. Moreover, neutrophil migration was evaluated with a Boyden chamber assay using zymosan-activated serum (C5a) as a chemoattractant.RESULTS: SFC dose-dependently inhibited the superoxide production induced by all the stimuli examined. Similarly, SFC-treatment inhibited the activity of NADPH oxidase in the membrane fraction of neutrophils. In addition, SFC suppressed chemotaxis toward zymosan-activated serum in a dose-dependent fashion.CONCLUSIONS: In this study, we revealed that SFC could suppress neutrophil functions such as superoxide generation (NADPH oxidase activity) and migration. During episodes of IBD, stimulated neutrophils are accumulated in inflamed mucosa via the migration through vascular endothelium, where they release ROS such as superoxide. Thus, the present observations suggest that SFC may have a potential to reduce mucosal injury by suppressing migration and ROS generation by neutrophils in UC.

目的:抗ヒトOrotate Phosphoribosyl Transferase(OPRT)抗体を用いたOPRT免疫組織化学染色とOPRT酵素活性,臨床病理学的背景因子およびOPRT翻訳領域にあるsingle nucleotide polymorphisms(SNPs)との関連性について検討した.対象:2003年12月から2004年7月までに当科で外科的切除された大腸癌35例とした.成績:OPRT活性は癌部が正常部に比べて有意に高値を示した(p

INTRODUCTION: Mesalazine(Pentasa) is widely used for ulcerative colitis(UC). However, the anti-inflammatory actions of Mesalazine are not fully understood. In this study, we evaluated the actions of Mesalazine in vitro on the functions of neutrophil, a major inflammatory cell. Furthermore, we evaluated the combined effects of Mesalazine and sodium ferrous citrate(SFC) on neutrophil functions, since SFC is clinically administered to UC patients together with Mesalazine for treatment of chronic anemia associated with UC. METHODS: Superoxide production by neutrophils was measured on the basis of superoxide dismutaseinhibitable cytochrome c reduction. Preincubated neutrophils were stimulated with fMLP, PMA or complement-opsonized zymosan particles. Cytochrome c reduction was calculated by the absorbance difference at 550 nm. Neutrophil migration was measured by a Boyden chamber assay with using zymosan-activated serum (C5a) as a chemotactic factor. Neutrophils were preincubated in the presence of 0.01-0.5mM Mesalazine and then incubated with fMLP. The cells were further incubated with phycoerythrin (PE)-labeled anti-CD11b monoclonal antibody, and the expression of CD11b was analyzed by flow cytometry. Neutrophils were preincubated with 5μg/ml SFC and 0.01-0.1mM Mesalazine, and superoxide generation and migration were measured.RESULTS: Mesalazine dose-dependently inhibited the superoxide production induced by PMA and fMLP at > 0.05 mM. Furthermore, Mesalazine inhibited chemotaxis toward zymosan-activated serum in a dosedependent fashion. Moreover, 0.5mM Mesalazine inhibited the fMLP-induced up-regulation of CD11b expression. Interestingly, Mesalazine combined with SFC inhibited the superoxide generation and migration by neutrophils, compared with Mesalazine alone. Thus, the present observations suggest that a combined administration of Mesalazine and SFC may have a potential to reduce mucosal injury by suppressing neutrophil functions (migration, ROS generation and expression of adhesion molecule) in UC.

Orotate phosphoribosyl transferase (OPRT) is the main enzyme that involves in phosphoribosylation of 5-fluorouracil (5-FU), an essential step that leads to tumor growth inhibition. In our study, the prognostic relevance of OPRT, thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) in resectable colorectal cancer (CRC) patients treated by oral 5-FU were compared to further clarify the prognostic value of OPRT. Tumor tissue was collected from 90 CRC patients and the patients were followed for 5.2 years (Median). TS, DPD and OPRT activities in the extract of tumor tissue were determined enzymatically. The cut-off value of OPRT (0.147 nmol/(min mg), TS (0.044 pmol/mg) and DPD (72.10 pmol/(min mg) were determined by maximal chi(2) method. Among these 5-FU metabolic enzymes, only high OPRT group demonstrated significantly better disease-free survival (DFS) (p = 0.0152) and better overall survival (p = 0.0078). In Cox regression analysis, node status (p < 0.0005) and OPRT (p = 0.044) were significant factors for DFS. OPRT activity in tumor tissue was a predictor of prognosis in resectable CRC patients treated by oral 5-FU-based adjuvant chemotherapy, and was useful to pick-up high risk patients independent from known prognosis factors.

Differentiation of naive CD4 T cells into Th2 cells is accompanied by chromatin remodeling including hyperacetylation of histones H3 and H4 in the nucleosomes associated with the IL-4, IL-13 and IL-5 genes. A conserved GATA3 response element (CGRE) containing four GATA consensus sequences was identified 1.6 kbp upstream of the IL-13 gene, corresponding with the 5′ border of the Th2-specific histone hyperacetylation region. The CGRE was shown to bind to GATA3, histone acetyl transferase complexes including CBP/p300, and RNA polymerase II. As for the IL-5 gene locus, CD28 costimulation selectively enhanced histone hyperacetylation through NF-κB activation and subsequent upregulation of GATA3. Chromatin remodeling of type 2 cytokine gene loci occurs also in developing Tc2 cells. IL-4 production and histone hyperacetylation in IL-4-associated nucleosomes in developing Tc2 cells were significantly lower than those of Th2 cells; however, cytokine production and histone hyperacetylation of IL-5 and IL-13 genes were equivalent. Developing Tc2 cells expressed lower GATA3 levels and dramatically increased levels of repressor of GATA (ROG). A ROG response element in the IL-13 gene exon 4 displayed Tc2-specific binding of ROG, HDAC1 and HDAC2. Thus, ROG may confer CD8 T cell-specific repression of histone hyperacetylation and activation of the IL-4 gene locus.

We have implemented a company-wide effort to progressively reduce the use of Pb(lead) and eventually eliminate this environmental pollutant from its products. As part of this effort, it has developed a new lead-free solder that consisting of Sn(tin), Zn(zinc), and Al(aluminum) and yet offers superior productivity and joint reliability. The new lead-free solder has a melting point equivalent to that of a SnPb eutectic solder, and enables devices to be packaged at a lower temperature than with the increasingly popular Sn(tin), Ag(silver), Cu(copper) solder. Thus, the new lead-free solder accelerates the elimination of Pb from products. we have already used printed circuit boards containing the new lead-free solder in some products, and plans to extend its use to other products. We further mounted SnZnAl solder balls onto Cu/Ni(nickel)/Au(gold) plated polyimide substrate at a joining temperature of 215 °C for CSP applications. It was confirmed that the joint interface between soldered ball and substrate at the initial stage was made of 2-layered compounds, i.e., AuZnSn on soldered side and ZnSnNi on substrate side. After 1000 h aging at 150 °C, the two layers compounds become one layer of ZnSnNiAu compound. No strength deterioration of Cu/Ni/Au/SnZnAl after the 1300 h shelf test was detected in the ball shear strength compared with the initial value. This paper describes the characteristics of the new lead-free solder and the results of a study on its solder ball CSP package.

High levels of foreign gene expression in mouse hepatocytes can be achieved by the rapid injection of a large volume of naked plasmid (pDNA) into animals via the tail vein, the so-called hydrodynamics-based procedure. In this study, we evaluated the efficacy of hydrodynamics-based tumor necrosis factor alpha (TNFalpha) transfer for tumor treatment, in which the naked pDNA encoding TNFalpha was administered into the tail vein following an intravenous injection of B16 melanoma cells. The mice treated with TNFalpha-expressing pDNA displayed a profound reduction in lung metastasis. These results suggest that the hydrodynamics-based transfer of naked pDNA is a convenient and efficient method of TNFalpha gene therapy against metastatic tumors.

神経鞘腫はSchwann細胞より発生する腫瘍であり,消化管,特に大腸より発生するものは稀である.症例は80歳,男性.検診で施行された注腸造影検査でS状結腸に20mm大の腫瘍を認めた.大腸内視鏡検査で腫瘍は粘膜下腫瘍であったため,腫瘍頂部の粘膜をEMRの手法で切除し,腫瘍深部の生検を試みたが確定診断を得ることはできなかった. GISTを疑い,腹腔鏡下S状結腸切除術を施行した.腫瘍は被膜に覆われ,粘膜筋板直下から筋層を貫き,漿膜下層まで及んでおり,紡錘形細胞が索状配列を呈していた.免疫染色ではc-kit陰性, S-100蛋白陽性, GFAP陽性を示し, S状結腸原発神経鞘腫と診断した.組織学的診断が得られない場合は, GISTなどのmalignant potentialを有する間葉系腫瘍も考慮し,診断的切除を選択することも必要と思われた.