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Nonsteroidal anti-inflammatory drugs (NSAIDs) induce apoptosis in a variety of cells, but the mechanism of this effect has not been fully elucidated. We report that diclofenac, a NSAID, induces growth inhibition and apoptosis of HL-60 cells through modulation of mitochondrial functions regulated by reactive oxygen species (ROS), Akt, caspase-8, and Bid. ROS generation occurs in an early stage of diclofenac-induced apoptosis preceding cytochrome c release, caspase activation, and DNA fragmentation. N-Acetyl-L-cysteine, an antioxidant, suppresses ROS generation, Akt inactivation, caspase-8 activation, and DNA fragmentation. Cyclic AMP, an inducer of Akt phosphorylation, suppresses Akt inactivation, Bid cleavage, and DNA fragmentation. LY294002, a PI3 kinase inhibitor, enhances Akt inactivation and DNA fragmentation. Ac-IETD-CHO, a caspase-8 inhibitor, suppresses Bid cleavage and DNA fragmentation. z-VAD-fmk, a universal caspase inhibitor, but not cyclosporin A (CsA), an inhibitor of mitochondrial membrane permeability transition, suppresses DNA fragmentation. These results suggest the sequential mechanism of diclofenac-induced apoptosis of HL-60 cells: ROS generation suppresses Akt activity, thereby activating caspase-8, which stimulates Bid cleavage and induces cytochrome c release and the activation of caspase-9 and-3 in a CsA-insensitive mechanism. Furthermore, we found that 2-methoxyestradiol (2-ME), a superoxide dismutase inhibitor, significantly enhances diclofenac-induced apoptosis; that is, diclofenac combined with 2-ME may have therapeutic potential in the treatment of human leukemia.

Although information about the mechanism of apoptosis is rapidly increasing, critical roles of mitochondria in the regulation of cell death remain to be elucidated. Recent studies revealed that reactive oxygen species (ROS) in cells increase prior to the onset of DNA fragmentation, a late phase event in apoptosis. An overload of Ca2+ induces a classic type of membrane permeability transition (CMPT) characterized by depolarization and swelling of mitochondria thereby releasing cytochrome c by a cyclosporin A-sensitive mechanism. In contrast, some agents induce apoptosis by increasing mitochondrial membrane permeability and swelling by some cyclosporin A- and Ca2+-insensitive mechanism; this phenomenon is defined as non-classic type of MPT (NCMPT). However, the role of ROS and antioxidants, such as α-tocopherol, in the mechanism of CMPT and NCMPT induction remains unclear. The present work overviews the role of ROS and α-tocopherol in the induction of CMPT and NCMPT.

Alpha-tocopherol (a form of vitamin E) is a fat-soluble vitamin that can prevent lipid peroxidation of cell membranes. This antioxidant activity of alpha-tocopherol can help to prevent cardiovascular disease, atherosclerosis and cancer. We investigated the alpha-tocopherol level and the expression of alpha-tocopherol transfer protein (alpha-TTP) in the leukocytes of children with leukemia. The plasma and erythrocyte alpha-tocopherol levels did not differ between children with leukemia and the control group. However, lymphocytes from children with leukemia had significantly lower alpha-tocopherol levels than lymphocytes from the controls (58.4 +/- 39.0 ng/mg protein versus 188.9 +/- 133.6, respectively; p < 0.05), despite the higher plasma alpha-tocopherol/cholesterol ratio in the leukemia group (5.83 +/- 1.64 micromol/mmol versus 4.34 +/- 0.96, respectively; p < 0.05). No significant differences in the plasma and leukocyte levels of isoprostanes (the oxidative metabolites of arachidonic acid) were seen between the leukemia patients and controls. The plasma level of acrolein, a marker of oxidative stress, was also similar in the two groups. Investigation of alpha-TTP expression by leukocytes using real-time PCR showed no difference between the two groups. These findings suggest that there may be comparable levels of lipid peroxidation in children with untreated leukemia and controls, despite the reduced alpha-tocopherol level in leukemic leukocytes.

Grain boundary diffusivity of 51 Cr and 59 Fe in high purity Fe-50 mass% Cr and Fe-60 mass% Cr alloys have been measured in the temperature range from 473 to 1273 K by use of a serial lathe sectioning technique. Below 653 K, the grain boundary diffusivity has been determined from the type C kinetics, whereas above 923 K it has been obtained from the type B kinetics. Linear Arrhenius lines have been observed for both types of B and C kinetics. Putting the grain boundary width δ at 5.10–10 m for the type B kinetics, the Arrhenius lines for the grain boundary self-diffusion coefficients Dgb in Fe-50 mass% Cr alloy obtained from both types kinetics are smoothly expressed as a single line. They can be expressed as :[math] for chromium and[math] for iron.The grain boundary diffusivities of 51 Cr and 59 Fe in Fe-60 mass% Cr alloy measured above 973 K are only slightly larger than those in Fe-50 mass% Cr alloy. The ratio of the activation energy for grain boundary diffusion to that for volume diffusion in the Fe-50 mass% Cr alloy is 0.35 which is much smaller than 0.6-0.7 observed in many metals and alloys.

The immune response caused by liposome stimulation was studied by assessing the level of several cytokines released from human peripheral blood cells. Liposome stimulation resulted in the release of IL-6, IL-10, IL-1beta, TNF-alpha and IFN-gamma. The size of the liposomes affected the degree of the cytokine releases with larger sized liposomes causing higher levels of cytokine induction. In addition, it appears that the lipid composition of liposomes had no effect on the degree of cytokine release. The release of cytokines occurred even in the absence of serum, suggesting that serum proteins did not contribute to liposome stimulation in peripheral blood cells. The release of cytokines induced by liposome stimulation was inhibited by the presence of either protein kinase-C (PKC) or protein tyrosine kinase (PTK) inhibitor, but not by the presence of an endocytosis inhibitor. This indicates that signal transduction via PKC or PTK is necessary, in order for human peripheral blood cells to release cytokines (IL-6, IL-10, IL-1beta, TNF-alpha and IFN-gamma) as the result of liposome stimulation. These quantitative data on the release of cytokines by liposomal stimulation provide useful information for the development of rational drug delivery systems and the safety of cytokine induction via the use of liposomes.

with monoblasts (AML-M5a subtype) with CD13 (99%), CD33 (97%), CD34 (99%), CD11c (54%), CD117 (91%), human leukocyte antigen (HLA)-DR (99%), CD4 (7%), CD14 (2%), and CD64 (1%). Chromosomal analysis showed the normal 46XX. The analysis of DNA revealed that blasts had the internal tandem duplication of the McDonough feline sarcoma viral oncogene (fms) like. We did not perform the mutation analysis of nucleophosmin 1 (NPM1) gene. Microscopic diagnosis revealed the blasts to be medium to large in size, and they had a pale cytoplasm with azurophilic granules. Since 10% of blasts had cuplike nuclear invaginations spanning over 25% of the nuclear diameter (see Figure 1), we considered that these blasts satisfied the criteria for AML-cuplike described by Kussick et al [1]. We did not perform electron microscopic observations of these blasts. Two courses of chemotherapy failed bone marrow blasts to attenuate less than 5% of all nuclear cells. The salvage chemotherapy brought about complete remission; thus, she underwent allo-peripheral blood stem cell transplantation (allo-PBSCT), wherein the donor was her HLA-matched brother. Although these treatments led to complete remission initially, the leukemia relapsed in the bone marrow 5 months after allo-PBSCT. Interestingly, 2% of blasts at relapse retained the cuplike nuclear

A 48-year-old man with no remarkable medical history presented with upper abdominal pain for approximately 1 month. He was diagnosed as having pancreatic carcinoma with liver and lung metastasis and complicating carcinomatous peritonitis. Despite chemotherapy, his performance status worsened, his appetite deteriorated, and his pain became intolerable. The patient opted to return home for palliative care, and his parents, aged over 70 years, supported this decision. Although corticosteroid and opiate administration was attempted to improve appetite loss and pain, oral administration became difficult over a short span of time. Thus, treatment was switched from oxycodone to a fentanyl patch for opioid rotation. We also prescribed risperidone for the treatment of delirium. The patient once opted for "respite hospitalization" at a general hospital to relieve his aged parents' fatigue, and thereafter, he finally died at home. When rapid disease progression is expected, not only should a fully equipped environment for patients be ensured but concern for their caregivers should also be considered. For this purpose, cooperation and communication among multidisciplinary medical staff is indispensable.

Bone metastasis from lung cancer accounts for approximately 30% of all metastatic bone tumors. The median survival time of patients with stage IV lung cancer with bone metastases is 5.5 months and that of patients without bone metastases is 7.5 months. Here, we report 3 cases of spinal cord paralysis. All cases were assessed according to the Tokuhashi score. As the predicted survival time of these patients wasor = 6 months, we opted for conservative treatment. We administered chemotherapy and radiation therapy, ensured symptom control, provided nursing care (prevention of decubitus, position changing, defecation control, rehabilitation, and mental health care), and coordinated home medical care. Patient management was mediated by a multidisciplinary medical team. However, all 3 patients were unable to return home and died in the hospital within 1-2 months after the onset of spinal cord paralysis. Spinal metastases can be expected not only in patients with lung cancer but also in patients with other types of carcinomas. Early diagnosis and treatment and accurate prognosis prediction are essential. Rapid responses and cooperation from experts are required, and increased awareness regarding spinal metastases among health professionals is essential.

We herein present a case of pediatric therapy-related myelodysplastic syndrome (t-MDS) with complex karyotype who was treated with azacitidine (AZA) for AML1-EVI1 fusion transcript as minimal residual disease after allogeneic hematopoietic stem cell transplantation (HSCT). The patient was started on AZA 41 days after the HSCT without having achieved complete remission. After 9 cycles of AZA, the AML1-EVI1 fusion transcript disappeared, and there was no manifestation of graft versus host disease during AZA treatment. Preemptive AZA treatment for minimal residual disease has an acceptable safety profile and appears to be an effective strategy for preventing or substantially delaying hematological relapse in pediatric patients with high-risk myelodysplastic syndrome after HSCT.

SummaryThis study correlates changes in neutrophilic activity and endothelial injury with markers of hemostatic activity following the infusion of increasing concentrations of E. coli organisms. It focuses on the hemostatic response as a marker of microvascular injury and uses the response to increasing concentrations of E. coli to refine our definition of disseminated intravascular coagulation (DIC) and distinguish between a compensated (non-overt DIC) and uncompensated (overt DIC) response. We observed that the global coagulation tests reflected activation of the hemostatic system in a dose dependent manner (overt DIC) in the early phases (T+2 to 6 h) of the response to increasing concentrations of E. coli, but that they failed to do so in the late phases (T+ 24 to 48 h). We observed that molecular markers, soluble thrombomodulin and elastase, unlike thrombin/antithrombin and plasmin/antiplasmin complexes, remained elevated out to T+24 to 48 h indicating endothelial injury that persists beyond the initial inflammatory insult in compensated as well as uncompensated DIC.

Aim: We report a case of X-linked agammaglobulinemia complicated with endobronchial tuberculosis. Methods: We observed the patient’s clinical course and analysed his data retrospectively. Results: Interestingly, the T-cell proliferation activity in this patient was intact, and the CD4-positive T cells produced interferon gamma. However, the result of the quantiferon alpha-2b test was negative. Conclusion: The findings of this case suggest that the quantiferon alpha-2b test may not be diagnostic for tuberculosis not only in patients with T-cell disorders but also in cases with B-cell deficiencies such as X-linked agammaglobulinemia.

Long-term peritoneal dialysis (PD) causes chronic peritoneal damage. Peritoneal mesothelial cells (PMCs) play an important role in peritoneal function. We investigated the possibility of cell therapy using the PMCs to prevent peritoneal damage in PD patients. We harvested human PMCs from the PD effluent of PD patients. The PMCs were separated based on morphological characteristics into epithelial-like (Epi) cells and fibroblast-like (Fib) cells by the limiting dilution method. We transplanted these cells into nude mice whose parietal and visceral peritoneum were scratched by mechanical scraping. The transplanted cells were detected at the parietal and visceral peritoneum. Compared with the positive control, the Epi cell therapy group showed very few adhesions and exhibited no thickening of the parietal and visceral peritoneum. However, the group with Fib cell therapy could not inhibit peritoneal adhesion and thickening. In addition, hepatocyte growth factor was expressed by the grafted Epi cells but not Fib cells. Fib cells expressed vascular endothelial growth factor stronger than Epi cells. These two types of cells from the same patient showed different characteristics and effects for cell therapy. These findings suggest that the PMCs from the PD patient showed different characteristics, such as Epi cells and Fib cells, and the selection of PMCs is important for cell therapy on the point of not only the direct cellular interactions but also cytokine secretion from the grafted cells. Furthermore, the differences in the morphological cell characteristics may influence their role in peritoneal regeneration.

Before hemodialysis (HD), plasma levels of tissue factor (TF), free-TF pathway inhibitor (TFPI) and thrombo modulin (TM) were significantly higher in patients with HD than in healthy volunteers. Plasma levels of (T-F) TFPI and plasmin plasmin inhibitor complex (PPIC) were significantly higher in patients with HD than in healthy volunteers. During HD, plasma levels of TF and (T-F) TFPI were not significantly increased, but plasma levels of total TFPI and free TFPI at 1 hour after and at the end of HD were significantly increased, compared with levels before start of HD. Plasma level of PPIC 1 hour after start of HD was significantly higher than before start of HD, and plasma levels of thrombin antithrombin com plex (TAT). PPIC. D-dimer. TM, and protein C (PC) at the end of HD were significantly higher than before start of HD. In patients with thrombosis complications, plasma TF levels were significantly higher than in patients without thrombotic com plications during HD. Plasma levels of PC were significantly lower in patients with thrombotic complications than in patients without thrombotic complications. There was no significant difference between both groups during HD in hemostatic pa rameters, with the exception of TF and PC. Hemostatic abnormalities existed in patients with HD; espe cially, increased TF and decreased PC might cause thrombotic complications.

人工物質最強の毒物であり，ニュース等でも数多く取り上げられているダイオキシンの分析方法について，1998年以降の技術動向を調査した。ダイオキシンの分析は，環境中の気体，液体，固体に吸着して存在しているダイオキシンを採取する方法やダイオキシン分析を妨害する共存物質を取り除く方法，極微量ダイオキシンを正確に測定する手段において通常の分析よりも難しい部分が多い。国の定めた方法（公定法）では極微量のダイオキシンを正確に測定することができるが，時間がかかる，高額，熟練した技術者が必要等問題も多い。必要最低限の感度および精度を保ちながら，公定法より低価格，短時間でできる分析法の研究概要を紹介する。

X-linked agammaglobulinemia (XLA, i.e. Bruton’s agammaglobulinemia [1]) is an X chromosomal recessive primary immunodeficiency syndrome, which is caused by insufficiency of antibody production. In 1993, BTK (Bruton’s tyrosine kinase) gene, the critical gene of XLA, was discovered in Xq22 by Tsukada et al. [2] and Vetrie et al. [3]. We report a family case with XLA, and describe the difference of clinical outcome between the proband and his brothers.

The cubic NiP 2 crystal was prepared by a mechanical milling technique. All-solid-state cells with the NiP 2 electrode and the Li 2 S–P 2 S 5 glass-ceramic electrolyte exhibited high reversible capacity of over 600 mAh g −1 for 10 cycles at room temperature. The NiP 2 electrode in solid-state cells underwent the conversion process as reported in the cells using a liquid electrolyte. Lithiated nickel phosphides were also prepared by milling. The composites of Li–Ni–P ternary crystal (Li 3.4 Ni 10.6 P 7 or Li 2 Ni 12 P 7 ) and Li 3 P were obtained and they were used as an electrode with lithium source in an all-solid-state cell. The cell with the electrode was initially charged and then discharged and retained cell capacity of about 600 mAh g −1 . Nickel phosphides are potential negative electrodes with high capacity in all-solid-state lithium rechargeable batteries with sulfide electrolyte.

All-trans retinoic acid (ATRA) is a metabolite of vitamin A that is used for molecular targeting therapy of acute promyelocytic leukemia (APL). Most patients with relapse after ATRA therapy show acquired resistance to ATRA, which seems to result from low plasma levels of the drug during therapy. This may be because of increased oxidative catabolism of ATRA or expression of the multidrugresistance gene product. Moreover, we have reported that impaired absorption can lead to lower plasma ATRA levels in patients with gastrointestinal complications [1]. Here, we describe the pharmacokinetics of ATRA in a patient with Crohn’s disease. A 13-year-old girl had been diagnosed with Crohn’s disease 9 months previously after standard endoscopic and histological evaluation. Treatment with steroids and 5-aminosalicyclic acid (5-ASA), as well as parenteral nutrition, led to a gradual improvement of her clinical state. She had no complications, so oral steroids therapy was tapered. Suddenly, she developed fever and blackish diarrhea, and was admitted with severe anemia. The hemoglobin was 6.5 g/dL, so red cell transfusion was performed. Bone marrow aspiration gave a diagnosis of APL. The reverse transcription-polymerase chain reaction detected a PML/RARa fusion chimeric gene. Colonoscopy revealed severe ulceration of the ascending colon and cecum.

ガス透過分離におけるガス透過の長期間の安定性と高効率化,更にガス透過効率の長期間の再現性を向上させた,新しいオンラインガス透過システムを構築した.このシステムでは新しく設計・製作したガス透過ユニットを組み込み,ユニット中を溶液が下方から上方に流れるようにし,気泡の発生を防ぐ工夫がほどこされ,更に測定後にガス透過ユニット内に滞留する溶液を排除するために2個の六方切り替えバルブを内蔵させている.ガスの安定的透過のために温度制御できる小型恒温槽を装着した.この装置中にガス透過ユニット,アンモニウムイオン定量用の反応コイルなどを組み入れ,精密測定の向上を目指した.ガス透過ユニットは,多孔質ポリテトラフルオロエチレン(PTFE)チューブとガラス管からなり,接続には樹脂製フェラル及びO-リングを用い,組み立てを容易にし,デッドボリュームも小さくした.ガス透過ユニット,反応コイル及び試料注入器などを恒温槽の中で一定温度に保つことにより,安定した再現性の良い測定が可能となった.ガス透過ユニット内に滞留する溶液を簡単かつ迅速に強制排除する機能も備えており,測定後はガス透過ユニット中の溶液を除いておくことにより,多孔質チューブの透過性能が長期間維持され,再活性化の操作をすることなく,高いガス透過効率を維持することができた.本システムを用いると,検量線は0～10,0～1.0ppmの範囲で良好な直線性を示し,1時間当たり30試料の分析が可能となった.実際に,河川水中のアンモニウムイオンの定量を行ったところ,インドフェノール誘導体/FIA/吸光光度法による定量値と良く一致し,試料に対する相対標準偏差は0.51,0.83%で,回収率は97～98%と良好であった.

試料採取現場での迅速な分析,すなわちオンサイト分析のための新しいポータブルフローインジェクション分析計の開発を行った.このEI装置では,160×160×320(mm)の箱の中にダブルプランジャーマイクロポンプ,試料注入器,検出器及び反応恒温槽を組み入れ,重量8kgと小型・軽量化を達成し,持ち運び容易な分析計とした.特に,525nmの発光ダイオード(LED)を光源とする新規検出器を開発することにより,小型,省電力化が可能となった.温度変化の大きな屋外での測定に耐えられるように,反応恒温槽も装備した.本FIA装置は交流100Vでのほか,直流12Vでも稼働し,カーバッテリーや市販のポータブルバッテリーを電源とするオンサイト分析に対応できるよう設計した.カーバッテリーを電源とした場合は長時間の連続運転が可能である。測定データはメモリーカードやRS232Cによりノート型パーソナルコンピュータへの通信も可能で,更に市販の表計算ソフトを利用してデータ処理が可能である.本装置を窒素酸化物のオンサイト分析に応用した.硝酸,亜硝酸イオン標準液による検量線は,0～1.0ppm,0～100ppbの範囲で直線性,再現性共に良好で,1時間当たり40～50試料の分析が可能であった.検出限界(S/N=3)は硝酸,亜硝酸態窒素として0.5ppbと通常のFIAシステムを用いる場合と全く変わらない優れた性能を有することが分かった.河川水中の硝酸亜硝酸イオンのオンサイト分析では10回の繰り返し測定における相対標準偏差は0.65%及び0.15%と良好な結果が得られた.また,血清中の一酸化窒素(NO)の酸化代謝物としての硝酸,亜硝酸イオン濃度のオンサイト分析を行い,良好な結果を得た.本研究で検討したポータブルバッテリーでは1回の充電で5時間の連続運転が可能であった.

Patients with a variety of nail disorders were treated with diode low reactive-level laser therapy (LLLT). Three representative cases are presented. Case 1 was a 28-year-old Japanese female with idiopathic twenty nail dystrophy. She was treated over one year with 79 days of LLLT. After LLLT, the rough nail surface on all the fingers improved remarkably. Case 2 was a 53-year-old Japanese female with green nails. She was treated over four months with 22 days of LLLT. The greenish color of the nail plates completely disappeared following LLLT. Case 3 was a 51-year-old Japanese male with longitudinal ridges, distal interphalangeal (DIP) joint arthritis and non-allergic drug eruptions. He was treated over about nineteen months with 94 LLLT sessions. After treatment with LLLT, the nail plate with longitudinal ridges became firm and longitudinal ridges slightly improved but his DIP arthritis improved remarkably. In all three representative cases, conventional treatments had failed to resolve the problems, which all responded well to LLLT. To date, a total of 22 patients with a variety of nail disorders have been successfully treated with GaAlAs diode LLLT. It is suggested that 830 nm GaAlAs diode LLLT should thus be added to the armamentarium of dermatologists and other appropriate specialists who see abnormal pathologies of the nails and associated conditions in the course of their practice.

All-trans retinoic acid (ATRA) is used as differentiation therapy for acute promyelocytic leukemia (APL). The two major adverse effects of ATRA therapy are hyperleukocytosis and retinoic acid syndrome. In order to prevent these adverse effects, low-dose ATRA therapy (25 mg/m 2 /d) has been tried in adults. Accordingly we assessed the pharmacokinetics of low-dose ATRA in children with cancer. Four children (one with APL and three with other advanced cancer) were administered ATRA and its pharmacokinetics were evaluated. In three patients, the pharmacokinetic parameters of ATRA were similar to those previously determined for APL patients in remission, but the values were lower in one patient. Low-dose ATRA was effective for APL, but not for the solid tumors. This therapy did not cause any severe toxicity. Further studies are needed to determine the optimum ATRA regimen and to evaluate low-dose ATRA combined with chemotherapy in children with APL.

ツムラ漢方製剤エキス顆粒 (医療用) の各方剤についてK含量を測定し, その全貌を明らかにし, 1包中のK含量は4.7-50.2mg (1g当1.88-16.52mg) の範囲にあることを確認した.128方剤のうち10mg未満のKを含むものは10品目, 10mg以上20mg未満のものは43品目, 20mg以上30mg未満のものは49品目, あわせて92品目 (71%) が10mg以上30mg未満の範囲にある. 30mg以上35mg未満のものは17品目, 35mg以上40mg未満のものは6品目, 40mg以上のものは大防風湯, 清上防風湯, 荊芥蓮翹湯の3品目となっている.1包中10mg未満のKを含有する方剤の場合は, 1日3回服用するとしてそのK量はせいぜい30mgであり, 米飯一杯のK量39mgと比較して特に問題とする量ではなかろう. 10mg以上30mg未満のKを含む方剤は1日の食餌により体内に入るK量に比べれば, それほど問題とするほどの量ではないが, 食事由来のKに漢方製剤由来のKが上積みされることを考慮して血漿K値を参照しながら使ってゆくべきであろう. 30mg以上のKを含有する26方剤については, さらに一層の注意が必要である.なお方剤中のK含量と, 方剤中に含まれる乾燥エキス量との間には正の相関が認められた.

Expression of retinoic acid receptor–target genes during retinoic acid therapy for acute promyelocytic leukemia

Necrotizing meningoencephalitis (NME), necrotizing leukoencephalitis (NLE) and granulomatous meningoencephalomyelitis (GME) are common idiopathic inflammatory central nervous system (CNS) diseases with unknown etiology in dogs. We previously showed that IgG autoantibodies in the cerebrospinal fluid (CSF) of NME cases reacted to unknown brain proteins as well as to glial fibrillary acidic protein (GFAP). In the present report, we evaluated the autoantibodies against transglutaminase2 (TG2) in the canine CNS diseases. CSF samples obtained from dogs with NME (n=19), NLE (n=7), GME (n=11) and miscellaneous CNS diseases (n=12) were subjected. CSFs from 20 healthy dogs were used as controls. Indirect fluorescent antibody test on the canine cerebrum revealed astrocyte-binding IgG in the CSF of NME. After absorption of the CSF with bovine GFAP, the CSF still possessed the reactivity to astrocytes. Double-color staining showed clear colocalization of the autoantibodies and anti-human TG2 rabbit polyclonal IgG. An immunoblot assay against human recombinant TG2 revealed anti-TG2 IgG in the CSF from dogs with NME, NLE and GME. The CSF of canine idiopathic encephalitis cases, notably of NME, tended to show high ELISA OD values against human recombinant TG2 compared to healthy controls. The presence of anti-TG2 autoantibodies in the CSF may contribute to the elucidation of the etiology of canine NME, NLE and GME.

We herein present the case of an acute monoblastic leukemia infant who relapsed with acute lymphoblastic leukemia (ALL). Complete remission was achieved after an acute myeloblastic leukemia-directed chemotherapy, but the patient relapsed with pro-B ALL. As he did not respond to acute myeloblastic leukemia-type reinduction therapy, he was switched to ALL-type chemotherapy, and a complete remission was achieved. Unrelated cord blood stem cell transplantation was performed, but he relapsed with pro-B ALL again. After he received ALL-type chemotherapy, he underwent another bone marrow transplant from his human leukocyte antigen mismatch mother, and has been free from recurrence for over 8 months.

BACKGROUND: Minimal residual disease (MRD) level after induction (Time Point1:TP1) and before consolidation therapy (Time Point2:TP2) has a strong impact in prediction of outcome for childhood acute lymphoblastic leukemia and it has clinical utility of a prognostic factor to stratify the risk groups in many current studies. We measured MRD levels on various time points to evaluate their prognostic significance in MRD-based augmented therapy. PATIENTS & METHODS: From June 2004 to September 2009, we prospectively assigned 333 consecutive children with ALL, 1〜19 years of age, to receive one of three treatment protocols on the stratification based on National Cancer Institute (NCI) risk criteria. NCI standard risk:SR, NCI high risk:HR and those with a white blood cell count≧1000x109 per L was defined as high-high risk:HHR. Patients were stratified again at TP2, patients with MRD level over 10-3 were assigned to salvage arm, treated in augmented therapy. Among 333 patients, 326 were eligible and 245 were MRD quantifiable. Then, we re-evaluated those samples by RQ-PCR according to the guideline of Euro MRD. Finally 167 cases were analyzed at 7 time points of MRD quantification in the CCLSG ALL2004 study. RESULTS: The overall 5-year event free survival (5-EFS) rate for ALL2004 was 82.5±2.1% (n=326), and 5-EFS of SR group (n=267), HR group (n=86) and HHR group (n=33) were 84.8±2.5%, 80.1±4.3% and 74.7±7.8%, respectively. In the SR group, 5-EFS of standard therapy group (n=124) with TP2 MRD TP1, at week 7, at the end of induction, 5-year relapse free survival (5-RFS) was 87.4.% in MRD negative cases (n=115) and 58.3% in MRD positive cases (n=36), and the differences were significant. TP2,week13, during consolidation, and TP3, at week 19 or 22, and at TP4, at week 26 to 28, had significant difference on 5-RFS with cut off of both 10-3 and 10-4. TP7, at the end of therapy, 2 patients were MRD positive and were thought molecular relapse. Additionally, we investigated changes in MRD levels of relapsed cases (n=35). MRD remained cases (n=4), MRD late disappeared cases (n=4), and MRD converted to positivity cases (n=4) were observed. CONCLUSIONS: The impact of predictive power of PCR MRD at TP1 and TP2 was validated, still more TP3 and TP4 were also statistically significant in CCLSG ALL2004 study. Even the patients whose final MRD level was negative could relapse, it is needed to investigate other prognostic factor except MRD. Disclosures Imai: Juno Therapeutics: Patents & Royalties.

This study was conducted to investigate the effect of geological succession of sediment on macrophyte and microbiota in aquifer ecosystem. The research field was selected as Yatsu Tidal Flat where located in urban coastal zone in Tokyo Inner Bay. This tidal flat has been registered under Wildlife Protection Area of Japan in 1988 and the Ramsar Convention in 1993, and just as the tidal flat that was left from reclamation of waterfront zone of urban area. It is very important to obtain some information about these ecosystem characteristics of such nature left in urban area, because creation of artificial tidal flat is recognized as one of important enterprises to construct urban ecosystem. In recent years, a large chlorophyceae, Ulva spp. (mainly Ulva pertusa and Ulva japonica) which is recognized as a biotic indicator of eutrophicated sea area, has become observed in Yatsu tidal flat, and it is considered that Ulva spp. give a large effect to water quality. Ulva spp. has high absorbent potential of nutrient salts at its growth phase, so this macrophyte is considered to place the microbiota such as zooplankton and phytoplankton under its control. In the first place, the geological succession as muddy to sandy occurred because of water quality change as fresh water to sea water. This water change resulted from the decrease of drainage with the sewer system service in basin area, and lead oligotrophication of the sediment. Ulva spp. is one of the biological indicater for sea water and sandy sediment and eutrophicated environment. Because of the irregular growth of Ulva spp., the quantity of phytoplankton decreased drastically, and the dominant species of phytoplankton and zooplankton transited from ideal to unsuitable food organisms for macrobenthos which is important food for the migratory birds, and the production potential of this aquifer ecosystem became in fragility.

We report a case of X-linked agammaglobulinemia complicated with endobronchial tuberculosis.We observed the patient's clinical course and analysed his data retrospectively.Interestingly, the T-cell proliferation activity in this patient was intact, and the CD4-positive T cells produced interferon gamma. However, the result of the quantiferon alpha-2b test was negative.The findings of this case suggest that the quantiferon alpha-2b test may not be diagnostic for tuberculosis not only in patients with T-cell disorders but also in cases with B-cell deficiencies such as X-linked agammaglobulinemia.

Recombinant canine interferon-γ (rCaIFN-γ) produced by a baculovirus expression system has therapeutic efficacy against atopic dermatitis in dogs. Although the mechanism of action of rCaIFN-γ is not completely understood, rCaIFN-γ is thought to downregulate the activity of interleukin-4- and interleukin-5-producing T helper 2 cells. However, rCaIFN-γ may also act directly on canine keratinocytes by inhibiting the release of inflammatory mediators. In this study, we investigated the effects of rCaIFN-γ on cytokine and chemokine mRNA transcription in a canine keratinocyte cell line, CPEK. It was found that granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA transcription was significantly inhibited after treatment with rCaIFN-γ (P0.001), whereas transforming growth factor-β and CC chemokine ligand 17 mRNA levels were unchanged. This study suggests that rCaIFN-γ may suppress GM-CSF production from canine keratinocytes, although further studies are required to confirm this.

A CC chemokine, CCL17/TARC, has been shown to be a factor in the immunopathogenesis of canine atopic dermatitis (cAD). In canine keratinocytes, the transcription of CCL17 mRNA is preferentially induced by tumor necrosis factor-alpha (TNF-α); however, its regulatory mechanism has not been elucidated. The aim of the present study is to clarify the regulatory mechanism of TNF-α-induced CCL17 mRNA transcription in canine keratinocytes leading to the development of a chemokine-targeted therapy for cAD. In a cell line of canine epidermal keratinocyte, CPEK, stimulation with TNF-α induced not only the activation of nuclear factor-kappa B (NF-κB) but also the phosphorylation of c-Jun-N-terminal kinase (JNK) and mitogen-activated protein kinase p38 (p38). Extracellular signal-regulated kinase (ERK) was found to be constitutively phosphorylated, which was temporarily augmented by TNF-α. Results of the inhibition assay indicated that the CCL17 mRNA transcription level was significantly decreased by p38 inhibitors but was not altered by either JNK or NF-κB inhibitors. Surprisingly, the ERK inhibitor increased the transcription level of CCL17 mRNA. Stimulation with epidermal growth factor (EGF), an ERK activator, suppressed the transcription of CCL17 mRNA. The present results suggest that TNF-α-induced CCL17 mRNA transcription in CPEK is positively regulated by p38 but negatively controlled by ERK.

Background The laminin (LN) γ2 chain expression has been linked to tumor invasion and prognosis. To provide a convenient clinical use, procedures that analyze LNγ2 expression by using the serum and/or urine of patients should be developed. Methods The serum concentrations of the N-terminal fragments of the LNγ2 chain in 73 patients with head and neck squamous cell carcinomas were measured by immunoassay. Results The concentrations of the LNγ2 fragments ranged between 14.5 and 324.2 ng/mL, and the normal upper limit was estimated to be 50 ng/mL. The LNγ2 fragment concentrations increased according to the T classification. The amount of elevated LNγ2 fragment concentrations decreased after the use of curative treatments. Three patients displayed a continuous increase of the concentrations and subsequently died of the diseases. Conclusions The serum concentrations of the LNγ2 fragments may prove useful in assessing the treatment results and clinical courses of patients with head and neck squamous cell carcinomas. © 2008 Wiley Periodicals, Inc. Head Neck, 2008

Alpha-tocopherol (a form of vitamin E) is a fat-soluble vitamin that can prevent lipid peroxidation of cell membranes. This antioxidant activity of alpha-tocopherol can help to prevent cardiovascular disease, atherosclerosis and cancer. We investigated the alpha-tocopherol level and the expression of alpha-tocopherol transfer protein (alpha-TTP) in the leukocytes of children with leukemia. The plasma and erythrocyte alpha-tocopherol levels did not differ between children with leukemia and the control group. However, lymphocytes from children with leukemia had significantly lower alpha-tocopherol levels than lymphocytes from the controls (58.4 +/- 39.0 ng/mg protein versus 188.9 +/- 133.6, respectively; p0.05), despite the higher plasma alpha-tocopherol/cholesterol ratio in the leukemia group (5.83 +/- 1.64 micromol/mmol versus 4.34 +/- 0.96, respectively; p0.05). No significant differences in the plasma and leukocyte levels of isoprostanes (the oxidative metabolites of arachidonic acid) were seen between the leukemia patients and controls. The plasma level of acrolein, a marker of oxidative stress, was also similar in the two groups. Investigation of alpha-TTP expression by leukocytes using real-time PCR showed no difference between the two groups. These findings suggest that there may be comparable levels of lipid peroxidation in children with untreated leukemia and controls, despite the reduced alpha-tocopherol level in leukemic leukocytes.

Changes of hemostatic markers in 226 patients with disseminated intravascular coagulation (DIC) and hematopoietic disorders were examined after treatment of DIC. The changes in prothrombin time (PT) ratio, fibrinogen, fibrin and fibrinogen degradation products (FDP), antithrombin, and protein C, thrombin-antithrombin complex (TAT), plasmin-plasmin inhibitor complex (PPIC), and soluble fibrin monomer complex (SFMC) in all patients with DIC were significant during the clinical course of DIC, but those of D-dimer, thrombomodulin (TM), tissue factor (TF), and tissue factor pathway inhibitor (TFPI) were not. Activated partial thromboplastin time (aPTT) and PT were significantly longer in the poor response group than in good response group. Plasma levels of FDP, TAT, PPIC, SFMC, TM, and DIC score were significantly higher in poor response group than in good response. Protein C and antithrombin levels were significantly lower in poor response group than in good response group. The changes of PT ratio, fibrinogen, FDP, DIC score, antithrombin, plasmin inhibitor, and protein C were significant in the good response group, but these levels were not significant in the poor response group. The changes in plasma TAT and SFMC levels were significant in the good response group but were not in poor response group. The changes in D-dimer, TM, TF, or TFPI were not significant in both groups. These findings suggest that anticoagulant agents should be administered at levels below TAT 40 ng/mL or SFMC 300 μg/mL in patients with DIC and hematopoietic disorders.

Bone marrow failure is often associated with skeletal defects, as seen in Fanconi anemia [1], Diamond-Blackfan anemia [2], or thrombocytopenia-absent radius syndrome [3]. Congenital amegakaryocytic thrombocytopenia with radio-ulnar synostosis has been reported to be related with HOXA11 mutation [4]. We present a boy with congenital bone marrow failure with radio-ulnar synostosis, but without HOXA11 mutation. A boy, born to non-consanguineous parents, was delivered via Cesarean section at 31 weeks’ gestation for hydrops fetalis resulting from severe anemia. He had severe anemia (hemoglobin 2.7 g/dl), severe neutropenia (white blood cells 3,220/ll with 1.5% neutrophils), and moderate thrombocytopenia (platelets 89,000/ll) at birth. Neutrophils continued to be nearly absent and platelet counts progressively decreased below 20,000/ll by 2 weeks of age. He had no hepatosplenomegaly, lymphadenopathy, or cafe-au-lait spot, but had hydrocele testicle, sensorineural hearing loss, and overlapping fingers without abnormalities of bone. Supination of his bilateral forearms was restricted and X-rays revealed bilateral proximal radio-ulnar synostosis (Fig. 1). Bone marrow aspiration showed aplasia without dysplasia. Chromosomal analysis revealed a 46, XY, inv(9)(p12q13) karyotype, which is considered to be normal variant without physiologic role. He lacked a family history of hematological or skeletal disorders. No excessive chromosomal breakage was detected by mitomycin C stimulation, and no mutation was found for the Shwachman–Bodian–Diamond syndrome (SBDS) gene. He had repeated bacterial and viral infections after birth, requiring antibiotics and prophylactic antifungal agents. He also needed repeated transfusions of red blood cells and platelets until 8 months of age when he underwent a 5/6 human leukocyte antigen (HLA)-matched unrelated bone marrow transplantation (BMT). Pretransplant conditioning was with fludarabine, cyclophosphamide, and thoracicabdominal irradiation (2 Gy). Prophylaxis for graft-versushost disease (GVHD) was short-term methotrexate and tacrolimus. Engraftment was prompt and grade 1 acute cutaneous GVHD was observed. By 20? days following BMT, he became independent of all transfusions. He has had normal hematological studies for 14 months after BMT. Limb formation is regulated by HOX genes during embryonic development. The most 50 members of the HOXA and HOXD clusters (HOXA9–HOXA13 and HOXD9–HOXD13) are particularly important in limb development [5]. HOX genes are also well known to be associated with hematopoiesis and leukemogenesis [6]. The effects of Hoxa10 and Hoxd11 were found to be on forearm development from studies of mutant mice [7, 8]. Radio-ulnar synostosis with congenital thrombocytopenia has been reported to be associated with HOXA11 mutation [4], which prompted us to examine HOX genes. However, we detected no mutations of HOXA11 and HOXD11. Of the three patients with radio-ulnar synostosis and thrombocytopenia reported by Thompson et al. [9], one H. Yoshida Y. Hashii T. Okuda S. Kusuki E. Sato H. Ohta (&) K. Ozono Department of Pediatrics, Osaka University Graduate School of Medicine, Yamadaoka 2-2, Suita 565-0871, Japan e-mail: ohta@ped.med.osaka-u.ac.jp

Changes of hemostatic parameters during percutaneous transluminal coronary angioplasty (PTCA) in 75 patients with chronic coronary artery disease were evaluated. Plasma levels of D-dimer, soluble fibrin monomer, plasmin-a2 antiplasmin inhibitor complex, and tissue factor (TF) were significantly increased in all patients with chronic coronary artery disease. The activity of antithrombin and protein C and the levels of protein C antigen were significantly decreased 1 hr after PTCA, but they returned to normal range 1 day after PTCA. There was no significant difference in the level of plasma APC-PCI complex before and 1 hr after PTCA. The plasma levels of D-dimer, soluble fibrin monomer, thrombomodulin, TF and PPIC were significantly decreased 1 hr, and the plasma levels of plasmin-a2 antiplasmin inhibitor complex 1 day after PTCA. These findings suggest that the decrease of protein C and antithrombin resulted in activation of the coagulation system. One hour after PTCA, the plasma levels of (total-free) TF pathway inhibitor (TFPI) were significantly decreased, but the plasma levels of total and free-TFPI were significantly increased, suggesting that comsumption of (total-free) TFPI occurs during PTCA. Overall, these findings suggest that the hypercoagulable state improves during PTCA and that transient decrease of antithrombin, protein C, (total-free) TFPI or plasmin-a2 antiplasmin inhibitor complex may cause restenosis of coronary artery. Am. J. Hematol. 61:238‐242, 1999. © 1999 Wiley-Liss, Inc.