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A novel culture system where cancer cells are grown in the presence of nanoclay in adherent plates allows preparation of malignant and dispersed spheroids compared with those formed under ultralow-adherent conditions.

Background In the process of community building, it is important to create a place for multigenerational exchanges. To promote multigenerational exchanges in regional locations, it is essential to clarify whether such exchanges are related to government infrastructure, regional characteristics, and social capital, and how these exchanges contribute to community building. Methods A cross-sectional questionnaire study was conducted with representatives from 455 Chiiki no Cha-no-Ma (literal translation “community living room,” and hereafter “Cha-no-Ma”) in Niigata City, Japan. Responses were received from 405 representatives (response rate: 89.0 %), and 401 agreed to participate (4 declined). The survey details included basic information (e.g., date each location was established, frequency of meetings, number of caretakers and participants, qualifications of the representative), activities reflecting local culture, a social capital scale, the effects of the Cha-no-Ma implemented by the representative (12 items), challenges for management (16 items), and the implementation of multigenerational exchanges. Results Most of the age groups that participated in the Cha-no-Ma were elderly, and multigenerational exchanges took place in 125 locations (31.5 %). Items that had a significant connection to the implementation of multigenerational exchanges were “Frequency of meetings” (p p = 0.026). Binomial logistic regression analysis indicated that a high frequency of meetings was associated with the implementation of multigenerational exchanges (Odds ratio = 3.839). There was a significantly higher ratio of implementation of multigenerational exchanges when the effects were a “connection with the region” (p = 0.006) and “conversations with different generations” (p = 0.004), and when the challenge was “no support from residents” (p = 0.002). Conclusions Cha-no-Ma participation is low among young people. The following ideas can be considered in order to increase multigenerational exchanges in regional locations. These exchanges may be promoted by increasing the frequency of meetings with qualified personnel and by adding activities that reflect local culture, such as festivals and making local foods. This community-based study clearly indicates that implementing multigenerational exchanges is an important activity for community building because it is related to connection within the community.

Background: In the process of community building, it is important to create a place for multigenerational exchanges. To consider the promotion of multigenerational exchanges in regional locations, it is important to clarify whether operational systems, regional characteristics, and social capital are related to the implementation of multigenerational exchanges and how these exchanges contribute to community building.Methods: The participants were representatives from 455 Chiiki no Cha-no-Ma (hereafter “Cha-no-Ma”), and the study was implemented in Niigata City, Japan. There were 405 responses (a response rate of 89.0%), with the analytical target being the 401 locations that agreed to participate in the study. A cross-sectional study based on a self-assessment questionnaire method was implemented, and questionnaires were returned directly to the researchers after completion. The survey details included basic information (e.g., date of establishment, frequency of meetings, number of managers and users, qualification of the representative, etc.); characteristic of regional activities; a Social Capital Scale; the effects of the Cha-no-Ma being implemented by the representative (12 items); challenges for management (16 items); and the implementation of multigenerational exchanges.Results: The items that had a significant connection to the implementation of multigenerational exchanges were “Frequency of meetings” (p < 0.001) and “Characteristic regional activities” (p = 0.026). With the relevant factors, a binomial logistic regression analysis was conducted, and the odds ratio for those with a high frequency of meetings was 3.839.There was a significantly higher ratio of the implementation of multigenerational exchanges when the effects were a “connection with the region” (p = 0.006) and “conversations with different generations” (p = 0.004). There was a significantly higher ratio of the implementation of multigenerational exchanges when the challenge was “no support from residents” (p = 0.002).Conclusions: To promote multigenerational exchanges in regional locations, meetings should be held at least once a week, with qualified personnel and the use of regional activities such as making local foods and festivals. It is clear from this community-based study that the implementation of multigenerational exchanges is an important activity for community building because it is related to connection within the community.

症例は65歳の男性で，精神疾患にて他院に入院中であったが，腹部激痛，ショック状態のため当院に搬送された．胸部単純X線検査，胸腹部CTにて横隔膜ヘルニアに伴う消化管穿孔，汎発性胸・腹膜炎の診断で緊急手術を施行した．左横隔膜後縁に約5 cmのヘルニア門を認め，胃体上部大彎に約2 cmの穿孔を認めた．胃部分切除，ヘルニア門の直接縫合閉鎖・ドレナージ術を行った．術後も敗血症性ショックの状態が持続したが徐々に回復し，術後第104病日に前医に転院となった．Bochdalek孔ヘルニアの成人例は比較的まれとされている．予後は一般に良好とされているが，脱出臓器の嵌頓・穿孔例では重篤化することがあるため，症状の有無にかかわらず早期に手術を行う必要があると考えられた．

症例は74歳の女性で，2007年2月心窩部痛を主訴に近医を受診し，腹部超音波検査で膵頭部近傍の嚢胞性腫瘤を指摘され当院を受診した．腹部CT，超音波，MRI検査で肝尾状葉に内部に造影効果を示す隔壁のある7 cm大の多房性の嚢胞性腫瘤を認め，一部に石灰化を伴っていた．肝粘液性嚢胞腺腫（mucinous cystadenoma；以下，MCA）と診断し，肝左葉・左尾状葉切除術を施行した．摘出標本は，5.5 cm大の多房性嚢胞性腫瘍であり，嚢胞内容は淡黄色透明の粘液であった．病理組織学的検査で嚢胞壁は粘液を有する円柱上皮で被覆され，一部に卵巣様間質に類似する部分を認めた．異型は認められず，肝MCAと診断した．尾状葉原発の肝MCAはまれな疾患で，本邦報告は3例のみである．嚢胞腺腫と嚢胞腺癌を術前に鑑別することは困難であり，肝MCAと診断しても経過観察はせず外科的切除が必要である．

症例は55歳，男性．2005年9月，S状結腸のポリープに対してポリペクトミーを施行した．病理組織学的診断は中分化腺癌，Type0-Isp（16×13×12mm）．SM（表層部から8,400um），ly0，v0，深部断端陽性（100μm（VM（+））であった．このため，同年11月，腹腔鏡補助下S状結腸切除術，D3郭清術を施行した．病理組織学的検査で癌遺残はなく，リンパ節転移も認めなかった．経過観察中の2007年5月，腹部超音波検査で肝S8に2cmの腫瘤を指摘された．画像検査所見から転移性肝癌と診断し，肝S8部分切除術を施行した．病理組織診断は，中分化腺癌で，大腸癌からの肝転移と診断した．本邦の報告では大腸SM癌の肝転移は1.2％とされ，稀である．さらに，リンパ節転移，脈管侵襲ともに陰性症例での肝転移は2例報告されるのみである．教室では1992年以降，手術を施行した大腸SM癌330例中，リンパ節転移，脈管侵襲ともに陰性の肝転移症例はこの症例のみであった．低リスクであっても画像診断などの定期的検査は必要であると考えられた．

Background/Aim Prostaglandin E 1 (PGE 1 ) has a wide-ranging effect on cytoprotection. Overproduction of heat shock protein 70 (HSP70) in the liver protects hepatocytes under various pathologic conditions. In this study, we examined the effect of a nontoxic HSP-inducer, PGE 1 , on acute liver failure after 95% hepatectomy in rats. Methods PGE 1 or vehicle was intravenously administered to rats 30 min before and during hepatectomy. Results Nine of 30 rats pretreated with PGE 1 survived, whereas all 20 rats pretreated with vehicle died within 96 h after operation. During the 24-h postoperative period, PGE 1 significantly suppressed the release of alanine aminotransferase and elevation of hyaluronic acid. Histological examination showed that the vacuolized hepatocytes and round hepatocytes with pyknotic nuclei are frequently seen in rats pretreated with vehicle, whereas active regeneration is seen in rats pretreated with PGE 1 . During the first 24 h after surgery, HSP70 induction was absent in the residual livers of vehicle-treated rats. In contrast, PGE 1 stimulated the HSP accumulation within 24 h, and viable hepatocytes contained abundant HSP70 in their nuclei. Conclusion Our results suggest that PGE 1 may prevent acute liver failure after massive hepatectomy, at least in part, by enhancing HSP70 production in the residual liver.

The transcription factor Gata1 is expressed in several hematopoietic lineages and plays essential roles in normal hematopoietic development during embryonic stages. The lethality of Gata1-null embryos has precluded determination of its role in adult erythropoiesis. Here we have examined the effects of Gata1 loss in adult erythropoiesis using conditional Gata1 knockout mice expressing either interferon- or tamoxifen-inducible Cre recombinase (Mx-Cre and Tx-Cre, respectively). Mx-Cre–mediated Gata1 recombination, although incomplete, resulted in maturation arrest of Gata1-null erythroid cells at the proerythroblast stage, thrombocytopenia, and excessive proliferation of megakaryocytes in the spleen. Tx-Cre–mediated Gata1 recombination resulted in depletion of the erythroid compartment in bone marrow and spleen. Formation of the early and late erythroid progenitors in bone marrow was significantly reduced in the absence of Gata1. Furthermore, on treatment with a hemolytic agent, these mice failed to activate a stress erythropoietic response, despite the rising erythropoietin levels. These results indicate that, in addition to the requirement of Gata1 in adult megakaryopoiesis, Gata1 is necessary for steady-state erythropoiesis and for erythroid expansion in response to anemia. Thus, ablation of Gata1 in adult mice results in a condition resembling aplastic crisis in human.

Background/Aims Well-organized turnover of the extracellular matrix is important in liver regeneration, which is regulated by the plasminogen activating system. The aim of this study was to investigate the role of plasminogen activator inhibitor-1 (PAI-1) after excessive hepatectomy and to ascertain whether angiotensin-converting enzyme (ACE) inhibitors, which are PAI-1 inhibitors as well, successfully improve the survival rate of rats that have undergone 95% partial hepatectomy (PHx). Methods Using liver tissues sampled after 90% or 95% PHx, the expression of PAI-1 mRNA was evaluated using reverse transcription polymerase chain reaction. Hepatic PAI-1 protein and urokinase-type plasminogen activator (uPA) levels were determined by enzyme-linked immunosorbent assay. Survival study and cytodynamic analysis by 5-bromo-2′-deoxyuridine staining were performed to evaluate the effects of ACE inhibition. Results The levels of PAI-1 mRNA and hepatic PAI-1 protein in the 95% PHx group peaked and were then maintained. By contrast, the uPA level fell relative to the 90% PHx group. Additionally, the hepatic PAI-1 protein level decreased and the survival rate improved in the 95% PHx rats that had undergone ACE inhibition. Conclusions Our experimental results suggest that PAI-1 plays a role in the occurrence of liver failure after excessive hepatectomy via accelerated maturation of pro-uPA and fibrinolytic factors. These are potential molecular therapeutic targets for liver failure after excessive hepatectomy.

Transcription factor GATA-1 is essential for erythroid cell differentiation. GATA-binding motifs have been found in the regulatory regions of various erythroid-specific genes, suggesting that GATA-1 contributes to gene regulation during the entire process of erythropoiesis. A GATA-1 germ-line mutation results in embryonic lethality due to defective primitive erythropoiesis and GATA-1-null embryonic stem cells fails to differentiate beyond the proerythroblast stage. Therefore, the precise roles of GATA-1 in the later stages of erythropoiesis could not be clarified. Under the control of a GATA-1 gene hematopoietic regulatory domain, a GATA-1 mutant lacking the N-finger domain (DeltaNF mutant) was over-expressed in mice. These mice exhibited abnormal morphology in peripheral red blood cells (RBCs), reticulocytosis, splenomegaly, and erythroid hyperplasia, indicating compensated hemolysis. These mice were extremely sensitive to phenylhydrazine (PHZ), an agent that induces hemolysis, and their RBCs were osmotically fragile. Importantly, the hemolytic response to PHZ was partially restored by the simultaneous expression of wild-type GATA-1 with the DeltaNF mutant, supporting our contention that DeltaNF protein competitively inhibits the function of endogenous GATA-1. These data provide the first in vivo evidence that the NF domain contributes to the gene regulation that is critical for differentiation and survival of mature RBCs in postnatal erythropoiesis.

Cancer cells undergo distinct metabolic changes to cope with their hypoxic environment. These changes are achieved at least partly by the action of transcriptional factors called hypoxia-inducible factors (HIFs). We investigated gene expression in cultured human colon cancer cells induced by hypoxic conditions with special reference to cell-adhesion molecules and carbohydrate determinants having cell-adhesive activity by using DNA-microarray and RT-PCR techniques. Hypoxic culture of colon cancer cells induced a marked increase in expression of selectin ligands, the sialyl Lewis x and sialyl Lewis a determinants at the cell surface, which led to a definite increase in cancer cell adhesion to endothelial E-selectin. The transcription of genes for fucosyltransferase VII ( FUT7 ), sialyltransferase ST3Gal-I ( ST3O ), and UDP-galactose transporter-1 ( UGT1 ), which are all known to be involved in the synthesis of the carbohydrate ligands for E-selectin, was significantly induced in cancer cells by hypoxic culture. In addition, a remarkable induction was detected in the genes for syndecan-4 ( SDC4 ) and α5-integrin ( ITGA5 ), the cell-adhesion molecules involved in the enhanced adhesion of cancer cells to fibronectin. The transcriptional induction by hypoxia was reproduced in the luciferase-reporter assays for these genes, which were significantly suppressed by the co-transfection of a dominant-negative form of HIF. These results indicate that the metabolic shifts of cancer cells partly mediated by HIFs significantly enhance their adhesion to vascular endothelial cells, through both selectin- and integrin-mediated pathways, and suggest that this enhancement further facilitates hematogenous metastasis of cancers and tumor angiogenesis.

Cellular biosynthesis of macromolecules often involves highly branched enzyme pathways, thus cellular regulation of such pathways could be rather difficult. To understand the regulatory mechanism, a systematic approach could be useful. We genetically analyzed a branched biosynthetic pathway for glycosphingolipid (GSL) GM1 using correlation index-based responsible enzyme gene screening (CIRES), a novel quantitative phenotype-genotype correlation analysis. CIRES utilizes transcriptomic profiles obtained from multiple cells. Among a panel of B cell lines, expression of GM1 was negatively correlated with and suppressed by gene expression of CD77 synthase (CD77Syn), whereas no significant positive correlation was found for enzymes actually biosynthesizing GM1. Unexpectedly, a GM1-suppressive phenotype was also observed in the expression of catalytically inactive CD77Syn, ruling out catalytic consumption of lactosylceramide (LacCer) as the main cause for such negative regulation. Rather, CD77Syn seemed to limit other branching reaction(s) by targeting LacCer synthase (LacCerSyn), a proximal enzyme in the pathway, because they were closely localized in the Golgi apparatus and formed a complex. Moreover, turnover of LacCerSyn was accelerated upon CD77Syn expression to globally change the GSL species expressed. Collectively, these data suggest that transcriptomic assessment of macromolecule biosynthetic pathways can disclose a global regulatory mechanism(s) even when unexpected.

Post-translational protein modifications are systems designed to expand restricted genomic information through functional conversion of target molecules. Ubiquitin-like post-translational modifiers regulate numerous cellular events through their covalent linkages to target protein(s) by an enzymatic cascade analogous to ubiquitylation consisting of E1 (activating), E2 (conjugating) and E3 (ligating) enzymes. In this study, we report the essential role of Uba5, a specific activating enzyme for the ubiquitin-like modifier, Ufm1, in erythroid development. Mice lacking Uba5 exhibited severe anaemia, followed by death in utero. Although Uba5 was dispensable for the production of erythropoietin, its genetic loss led to impaired development of megakaryocyte and erythroid progenitors from common myeloid progenitors. Intriguingly, transgenic expression of Uba5 in the erythroid lineage rescued the Uba5-deficient embryos from anaemia and prolonged their survival, demonstrating the importance of Uba5 in cell-autonomous erythroid differentiation. Our results suggest that one of the ubiquitin-like protein modification systems, the Ufm1 system, is involved in the regulation of haematopoiesis., Post-translational modifications are important in regulating protein function and turnover, and Ufm1 is part of a recently identified protein modification system. In this study, the authors show that Uba5, a component of the Ufm1 system, is important for regulating haematopoiesis and the differentiation of erythroid cells.

The anti-CD20 monoclonal antibody (Ab) rituximab is accepted to be an effective therapeutic Ab for malignant B-cell lymphoma; however, discovery of other cell surface antigens is required for the option of antibody medicine. Considering that many tumor-associated antigens are glycans, we have searched glycoconjugates for the candidate antigens that therapeutic Abs target. To this end, we first focused on the difference in the glycogenes expression in terms of Epstein-Barr virus (EBV) infection of a Burkitt's lymphoma cell line, Akata. Using DNA array, flow cytometry and Western blotting, we found that Thy1 was highly expressed in EBV-positive Akata cells. Subsequently, Thy1 was found to be expressed in other B-cell lymphoma cell lines: BJAB, MutuI, and MutuIII, irrespective of EBV infection. Treatment of these cells with an anti-Thy1 monoclonal antibody inhibited proliferation more strongly than the therapeutic Ab rituximab. The B-cell lymphoma cell lines were classified based on the extent of the proliferation inhibition, which was not correlated with the expression level of Thy1. It is suggested that stable residence of receptor tyrosine kinases in lipid rafts sustains cell growth in B-cell lymphoma cells.

Allergic reactions to oxaliplatin can be severe and are an important cause of discontinuation of treatment. A retrospective review was performed for 105 patients who received FOLFOX regimens between May 2005 and June 2007. Twenty-five cases (23.8%) of allergic reactions were identified, including 9 late onset reactions (8.6%) and 16 immediate reactions (15.2%). Severe allergy (Grades 3 and 4) occurred in seven patients (6.7%). Re-introduction of FOLFOX was attempted for seven immediate onset patients with a severity grade of 1 or 2, and three of these patients (42.9%) showed relapse of allergy. In approximately 10% of the patients, FOLFOX had to be discontinued due to allergy before the disease became refractory to the regimen. Our experience indicates that allergy to oxaliplatin may be a significant concern and that methods are required for suppression of this allergy.

DNA microarray analysis enables us to provide a considerable amount of information on glyco-chain expression (Brazma et al. 2001; Ide et al. 2006). DNA microarray experiments are developed to obtain overall transcriptome of cells using DNAs spotted on a slide glass as probes and fluorescently labeled DNAs derived from mRNA samples. DNA microarray experiments require high-quality RNA. Thus, it is advisable to prepare RNA as intact as possible. The quality of RNA is usually assessed by the intactness of the peaks and the lack of degradation of RNA by capillary electrophoresis. This step is essential for the reliable final readout of microarray experiments. Following is the normal procedure operated in our laboratory for the DNA microarray experiments using RIKEN human and mouse glyco-chain-related DNA microarrays version 2 (human version: GEO platform GPL 3465 http://www.ncbi.nlm.nih.gov/geo/), which were designed by us and spotted by Takara Bio Inc.

The galactosylsphingosine psychosine (Psy) is one of the sphingolipids and induce the formation of multinuclear cells in several cell lines by inhibiting cytokinesis. In the present report, we show that intracellular organelles, including wheat germ agglutinin (WGA)-positive vesicles and early endosomes, are selectively dispersed by Psy. WGA is a conventional Golgi marker and WGA-positive vesicles appeared to co-localize with the Golgi apparatus in untreated cells. Psy treatment induced the dispersal of WGA-positive vesicles without affecting the structure of the Golgi apparatus, resulting in discrimination of WGA-positive vesicles from the Golgi apparatus. In sharp contrast to this effect of Psy, WGA-positive vesicles were not affected by brefeldin A treatment, which induced the disappearance of the Golgi apparatus. Immunostaining with anti-TGN46 antibodies revealed that a large portion of the WGA-positive vesicles were derived from the trans-Golgi network. Notably, the dispersed WGA-positive vesicles did not stain with anti-syntaxin 6, another marker of the trans-Golgi network. During cytokinesis, WGA-positive vesicles in the cytoplasm decreased, and WGA staining accumulated at the cleavage furrow, which was apparently inhibited by the presence of Psy. These data suggest that the transport of WGA-positive vesicles to the cleavage furrow is associated with the progression of cytokinesis.

S It remains unknown whether the normal adult liver contains bipotent stem/progenitor cells, and if it does, then what are the circumstances under which they proliferate. The aim of this study was to clarify whether the normal adult liver contains hepatic stem/progenitor cells, and if it does, will they be activated by extensive hepatectomy? Adult rat liver cells were isolated and cultured at a low-density, and the colony-forming assay was performed to evaluate the cell proliferative capacity. Immunocytochemistry and reverse transcription-polymerase chain reaction were used to investigate the multilineage differentiation capability. The rate of colony formation by cells from the normal liver and those from the regenerating liver after partial hepatectomy (PH) were compared to determine whether progenitor cell proliferation might be activated by PH. Only a few epithelial colonies (0.043+/-0.009% of nonparenchymal cells) continued to proliferate for more than 1 month. Reverse transcription-polymerase chain reaction and immnocytochemistry showed that these progenitor colonies expressed both hepatocyte and cholangiocyte markers. The proportion of progenitor cells that formed bipotential colonies did not differ significantly between the cells obtained from the normal and PH livers. Adult normal liver contains bipotent hepatic progenitor cells, but they are scarcely activated even after extensive hepatectomy.

Sialic acid (Sia) is a family of acidic nine-carbon sugars that occupies the nonreducing terminus of glycan chains. Diversity of Sia is achieved by variation in the linkage to the underlying sugar and modification of the Sia molecule. Here we identified Sia-dependent epitope specificity for GL7, a rat monoclonal antibody, to probe germinal centers upon T cell-dependent immunity. GL7 recognizes sialylated glycan(s), the alpha2,6-linked N-acetylneuraminic acid (Neu5Ac) on a lactosamine glycan chain(s), in both Sia modification- and Sia linkage-dependent manners. In mouse germinal center B cells, the expression of the GL7 epitope was upregulated due to the in situ repression of CMP-Neu5Ac hydroxylase (Cmah), the enzyme responsible for Sia modification of Neu5Ac to Neu5Gc. Such Cmah repression caused activation-dependent dynamic reduction of CD22 ligand expression without losing alpha2,6-linked sialylation in germinal centers. The in vivo function of Cmah was analyzed using gene-disrupted mice. Phenotypic analyses showed that Neu5Gc glycan functions as a negative regulator for B-cell activation in assays of T-cell-independent immunization response and splenic B-cell proliferation. Thus, Neu5Gc is required for optimal negative regulation, and the reaction is specifically suppressed in activated B cells, i.e., germinal center B cells.

Background Glycan biosynthesis occurs though a multi-step process that requires a variety of enzymes ranging from glycosyltransferases to those involved in cytosolic sugar metabolism. In many cases, glycan biosynthesis follows a glycan-specific, linear pathway. As glycosyltransferases are generally regulated at the level of transcription, assessing the overall transcriptional profile for glycan biosynthesis genes seems warranted. However, a systematic approach for assessing the correlation between glycan expression and glycan-related gene expression has not been reported previously. Methodology To facilitate genetic analysis of glycan biosynthesis, we sought to correlate the expression of genes involved in cell-surface glycan formation with the expression of the glycans, as detected by glycan-recognizing probes. We performed cross-sample comparisons of gene expression profiles using a newly developed, glycan-focused cDNA microarray. Cell-surface glycan expression profiles were obtained using flow cytometry of cells stained with plant lectins. Pearson's correlation coefficients were calculated for these profiles and were used to identify enzyme genes correlated with glycan biosynthesis. Conclusions This method, designated correlation index-based responsible-enzyme gene screening (CIRES), successfully identified genes already known to be involved in the biosynthesis of certain glycans. Our evaluation of CIRES indicates that it is useful for identifying genes involved in the biosynthesis of glycan chains that can be probed with lectins using flow cytometry.

Transcription factor GATA-1 is essential for erythroid cell differentiation. GATA-binding motifs have been found in the regulatory regions of various erythroid-specific genes, suggesting that GATA-1 contributes to gene regulation during the entire process of erythropoiesis. A GATA-1 germ-line mutation results in embryonic lethality due to defective primitive erythropoiesis and GATA-1-null embryonic stem cells fails to differentiate beyond the proerythroblast stage. Therefore, the precise roles of GATA-1 in the later stages of erythropoiesis could not be clarified. Under the control of a GATA-1 gene hematopoietic regulatory domain, a GATA-1 mutant lacking the N-finger domain (DeltaNF mutant) was over-expressed in mice. These mice exhibited abnormal morphology in peripheral red blood cells (RBCs), reticulocytosis, splenomegaly, and erythroid hyperplasia, indicating compensated hemolysis. These mice were extremely sensitive to phenylhydrazine (PHZ), an agent that induces hemolysis, and their RBCs were osmotically fragile. Importantly, the hemolytic response to PHZ was partially restored by the simultaneous expression of wild-type GATA-1 with the DeltaNF mutant, supporting our contention that DeltaNF protein competitively inhibits the function of endogenous GATA-1. These data provide the first in vivo evidence that the NF domain contributes to the gene regulation that is critical for differentiation and survival of mature RBCs in postnatal erythropoiesis.

Mannan-binding protein (MBP) is a Ca2+-dependent mammalian lectin that plays an important role in innate immunity. In this study, we found that ligand-bound MBP stimulates polymorphonuclear leukocytes (PMN) to induce cell aggregation and superoxide production. The biological response of PMN to ligand-bound MBP was dose- and time-dependent. The PMN aggregation and superoxide production induced by ligand-bound MBP was blocked completely by pertussis toxin, and partially blocked by a platelet activation factor receptor antagonist, TCV-309. These findings suggest that the ligand-bound MBP stimulates PMN through a putative MBP receptor(s) on PMN. Published in 2004.

We performed hepatic arterial infusion (HAI) chemotherapy for 4 patients with advanced gastric cancer who had undergone curative resection except for liver metastasis. The main antineoplastic drugs were 5-fluorouracil (5-FU), mitomycin C (MMC) and cisplatin (CDDP). A catheter was inserted into the hepatic artery by interventional radiological techniques in 3 patients and operatively in 1 patient. The response rate for 4 patients was 75% (CR2, PR1, PD1). The adverse events were Grade 3/4 nausea and/or vomiting in 2 cases. The HAI chemotherapy was effective and useful for patients with advanced gastric cancer who had no unresectable lesions except for liver metastasis.

In this review, we focus on sphingolipids as potential regulators of the induction of multinuclear cell formation through the inhibition of cytokinesis. A sphingolipid, psychosine (Psy) (galactosylsphingosine), was demonstrated to be a trigger lipid for the inhibition of cytokinesis and the induction of multinuclear giant cells associated with a sphingolipid metabolic disease, globoid cell leukodystrophy (GLD). Indeed, Psy is known to accumulate in the patients' brains. Interestingly, inhibition of sphingolipid biosynthesis also induced multinuclear cells. When cells were treated with a new immunosuppressant, ISP-1/myriocin, which inhibits serine palmitoyltransferase, the first step enzyme of sphingolipid biosynthesis, the cells underwent multinucleation and apoptosis. At present, a definitive model of the function of sphingolipids as to the induction of multinuclear cell formation is not available due to the rudimentary information but possible mechanisms are discussed.

Equine kidney and spleen contain a Forssman active glycosphingolipid, and the structure of this glycolipid has been reported to be that of a globopentaosylceramide (GalNAcalpha-1,3GalNAcbeta-1,3Galalpha-1, 4Galbeta-1,4Glcbeta-1,1'Ceramide). We found that equine kidney contains several other anti-Forssman antibody-reactive glycosphingolipids. One of these acidic Forssman active glycosphingolipids was isolated and characterized by means of NMR, mass spectrometry, permethylation studies, and TLC-immunostaining. This glycolipid contains three moles of galactose, one mole of glucose, three moles of N-acetylgalactosamine, one mole of N-acetylglucosamine, and one mole of N-acetylneuraminic acid, and is stained on TLC with anti-Forssman antibodies and anti-GM2 ganglioside antibodies. HOHAHA and ROESY experiments and permethylation studies showed this glycolipid oligosaccharide to be branched at the innermost galactose; one chain has an isoglobo structure with a terminal Forssman disaccharide and the other chain is branched through the linkage of N-acetylglucosaminebeta-1,6 to the inner galactose. The nonreducing end of the GM2 trisaccharide is linked to this glucosamine. The structure of the oligosaccharide of the glycolipid presented here is a novel type, having branched isoglobo-, ganglio-, and neolacto-series oligosaccharides. Mass spectrometric analyses indicated the ceramide moiety of the glycolipid to be composed predominantly of hydroxy fatty acids (C20:0, C22:0, C23:0, C24:0, and C25:0) and hydroxysphinganine. GalNAcalpha-1,3GalNAcbeta-1,3Galalpha-1,3[GalNAcbet a-1, 4(NeuAcalpha-2,3)Galbeta-1,4GlcNAcbeta-1,6]Galbeta+ ++-1,4Glcbeta-1, 1'Ceramide

A bone marrow proteoglycan (BMPG) has been purified which consists of the same amino acid sequence as that of pro-MBP, and produced two anti-BMPG mAbs. Serum levels of major basic protein (MBP), a cationic protein rich in the eosinophil granule, were measured in patients with eosinophilia or allergic diseases by an enzyme-linked immunosorbent assay (ELISA) using these mAbs. The serum levels of MBP in patients with eosinophilia (n = 64) and in those with allergic diseases without eosinophilia (n = 32) were elevated significantly (P < 0.001 and P = 0.038, respectively). There was a weak positive correlation between the serum levels of MBP and the eosinophil counts in the patients with eosinophilia (r = 0.38). Among these patients, extremely high serum levels of MBP were found in those with hypereosinophilic syndrome (HES) and Kimura's disease. Serum levels of MBP decreased more slowly than the eosinophil counts in patients with eosinophilia when treated by glucocorticoids. We conclude that measurement of serum levels of MBP is useful in evaluating the total-body proliferation and infiltration of eosinophils more accurately than following-up the eosinophil counts alone.

Superoxide dismutase (SOD) was covalently conjugated with the copolymer of divinyl ether and maleic anhydride (DIVEMA) with average molecular weight of 30000 and a narrow molecular weight distribution (Mw/Mn= 1.4). To avoid the crosslinkage and the decrease in enzymatic activity, a pHreversible amino group-protecting agent, 1,1-dimethylmaleic anhydride (DMMAn), was applied in the synthesis. The amino groups of SOD were firstly protected by DMMAn; and then, after the remaining amino groups were reacted with the acid anhydride groups of DIVEMA, the protective agent was removed by lowering of the pH to 6.0. By use of this protective agent the crosslinked product could be minimized. The DIVEMA-SOD conjugate could be purified by preparative GPC. The resulting DIVEMA-SOD conjugate, free from native SOD, retained almost the same enzymatic activity as that of native SOD as determined by the method of McCord.

An unusual case of chronic neutrophilic leukemia associated with paraproteinemia (IgG Type lambda and IgA Type K) was followed for 3 years. Clinical and hematological findings obtained during the entire course fulfil the previously reported criteria for the diagnosis of chronic neutrophilic leukemia. A marked infiltration of mature neutrophils in the spleen and liver was confirmed by postmortem examination.

長期間にわたる多量の常飲者で,過飲による増悪と,断酒による軽快を繰り返した2症例につき,2年以上経過を追跡して,経時的に腹腔鏡検査と肝生検を繰り返し検索した.アルコール性肝炎では,その繰り返しにより約2年間で壊死後性肝硬変が完成され,門脈性肝硬変では,アルコール性肝炎が繰り返されると約1年間で壊死後性肝硬変へと進展した.これらの観察から,アルコール性肝硬変の進展にはアルコール性肝炎が重大な影響を及ぼしていることが明らかにされた.