Immunotoxins are fusion proteins consisting of two elements, a targeting and a toxin moiety, and are designed for specific elimination of tumor cells. Previously we expressed a recombinant fusion protein consisting of the toxic fragment of Shiga toxin (A1) and GMCSF (A1-GMCSF) in Escherichia coli, and evaluated its cytotoxic properties in acute myeloid leukemia and colon carcinoma cell lines. In view of the specific cytotoxic effects of this immunotoxin, further detailed in-vitro and preclinical studies were undertaken. Large amounts of the recombinant protein of high purity and free of unwanted side products, such as lipopolysaccharides (LPS), were required. Since GMCSF is of mammalian origin and it requires proper disulfide bond formation, we intended to use the baculovirus expression vector system (BEVS) for the expression of the recombinant fusion protein. However, despite previous reports on the expression of several other immunotoxins by this system, the A1 derived fusion proteins revealed an inhibitory effect on baculoviral particle formation and even caused cell death in insect cells. This observation was further pursued and confirmed by the use of other baculoviral specific promoters. The salient features of this finding are described below.

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The academic ability of freshman widely spreads due to the variety of learning experience at high school. To respond to this problem, a basic mathematics and physics course was newly offered as an introductory engineering education at KIT in 2004. In order to improve the academic ability of mathematics and physics, various learning support programs are being carried out in this course. The questionnaire and interview of the student were executed in these programs. In these results, the learning experience at high school and the consciousness for the mathematics and physics of the student were verified.

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生物経済モデルによって豚の諸形質の経済価(EV)を推定するとともに,わが国の豚の育種構造での各々の種畜から産肉および繁殖形質に発現した累積割引総量(CDE)をgene flow法から求めた.形質1単位当りの売上収入は,熊本県内の屠畜場で1991年に得られた1,258頭の肉豚の枝肉情報の重回帰分析から推定した.ただし,肉豚1頭当りの収入は,1993年の大阪市場の格付け毎の年平均枝肉単価から算出した.
主な豚の選抜諸形質のEV(円•年^-1•頭^-1)は,肥育豚の発育率:16(g^-1),背脂肪の厚さ:-331(mm^-1),一腹当り正常産子数:764(頭^-1),離乳時事故率:-88(%^-1)および肥育豚事故率:-179(%^-1)であった.また,評価期間20年,割引率6.5%の時の,雌系統A,雌系統Bおよび雄系統Cから後代への遺伝子経路を経て,産肉形質および繁殖形質に発現したCDE(頭•半年•頭^-1)は,A母からはそれぞれ0.516,0.979,B父からはそれぞれ0.499,0.849またC父からはそれぞれ1.109,0.019であった.
重回帰分析と生産モデルを組み合せた本方法は,形質の利益に対する非線形性を考慮でき,さまざまな形質のEVの推定に有効と思われる.また,CDEによって補正されたEVを用いることで,経済的視点に立った豚の改良方向の決定と,将来の育種環境の不確実性を考慮した育種計画が可能となった.

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Ankle-foot orthosis (AFO)is an important aid for patients with gait disorder. We have investigated the relationship between hemiplegic ankle joints function and AFO joint characteristics during gait, because mechanical characteristics of AFO joint directly affect the walking function. In this article, we examined an ankle joint device using magneto-rheological fluid that rheological characteristics can be adjusted by changing intensity of magnetic field applied. The device is composed of a rotary cylinder and a short tube with restrictor connecting both cylinder ports, and its closed hydraulic circuit is filled with MR fluid. The moment and angular velocity relationship was measured for the device with a permanent magnet, and the resistant moment was evaluated under different magnet distance from the tube. This device was mounted on an experimental CFRP upright AFO accompanied with a magnet position adjustment mechanism. Gait of a hemiplegic volunteer with this AFO was improved in some time-distance parameters when viscosity was switched to low at initial stance phase and to high immediately before swing phase.

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The objective of this research was to develop a wireless system enabling communication between a Central Control Unit and four robots that worked in a model greenhouse. The wireless system could be applied to control multiple robots in fruit picking, pesticide spraying, and resource transportation. Two of the robots were designated as “work-robots”, traveling in work ways between plants. The other two “assist-robots”, moving in two passage ways, had the task of carrying the work-robots to designated work ways. The results showed that the success rate of system operations under varying conditions was consistently above 95%. The developed wireless control system has shown potential for application in real life greenhouse operations and as an educational tool.

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Background:　When multipass transmembrane molecules are located on the cell surface, there may be interaction with not only bioactive molecules but also pathogenic molecules in areas protruding outside the cell. In antibody-mediated autoimmune disorders, it has been found that the autoantibodies occasionally attack membrane molecules on the cell surface, thus causing the disease such as myasthenia gravis. In such cases, highly sensitive autoantibody detection technology is required for early diagnosis. However, autoantibody analysis technology that is specialized for membrane molecules is still under development. Here we demonstrate a novel method for detecting of antibodies against the extracellular portions of multipass transmembrane molecules. Methods:　Antibodies for muscarinic acetylcholine receptor type3 (M3R) were detected with two kinds of luciferase immunoprecipitaion systems (LIPS), conventional LIPS (cLIPS) and its modified application, termed modified LIPS (mLIPS). In mLIPS, antibodies against extracellular portions of membrane molecules could be preferentially detected. Results:　An antibody to the amino-terminal portion of human M3R was detected with modified LIPS with a high sensitivity. In contrast, an antibody to the carboxyl-terminal portion was not detected with mLIPS, because it did not interact with intracellular portions of M3R in living cells. We also found antibodies for M3R in a patient serum with Sjögren’s syndrome. Conclusion:　Our technology has a promising future, and we hope that it will be applied in the analysis of antibodies against a diverse range of multipass transmembrane molecules, including GPCRs.

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This paper is about a new efficient method for the implementation of a Block Proportionate Normalized Least Mean Square (BPNLMS++) adaptive filter using the Fermat Number Transform (FNT) and its inverse (IFNT). These transforms present advantages compared to Fast Fourier Transform (FFT) and the inverse (IFFT). An efficient state space method for implementing the FNT over rectangular windows is used in the cases where there is a large overlap between the consecutive input signals. This is called Generalized Sliding Fermat Number Transform (GSFNT) and is useful for reducing the computational complexity of finite ring convolvers and correlators. In this contribution, we propose, as a first objective, an efficient state algorithm with the purpose of reducing the complexity of IFNT. This algorithm, called Inverse Generalized Sliding Fermat Number Transform (IGSFNT), uses the technique of Generalized Sliding associated to matricial calculation in the Galois Field. The second objective is to realize an implementation of the BPNLMS++ adaptive filter using GSFNT and IGSFNT, which can significantly reduce the computation complexity of the filter implantation on digital signal processors.

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We studied the effect of a high glucose level on gap junctional intercellular communication (GJIC) activity in cultured vascular smooth muscle cells (VSMCs) using the fluorescent dye transfer method. After a 48-h incubation with 22mmol/l glucose (high glucose level), GJIC activity of VSMCs was significantly reduced as compared to the incubation with 5.5 mmol/l glucose (normal glucose level). Treatment of the cells with 12-Otetradecanoylphorbol-13-acetate (TPA;5×10^-8 mol/l), a protein kinase C (PKC) activator, for 1h also significantly reduced GJIC activity. In addition, treatment with calphostin C, a specific PKC inhibitor, for 3h completely restored the glucose-induced reduction of GJIC activity. Western blot analysis showed that connexin 43 (Cx43), which is the major functional protein of GJIC, is present in multiphosphorylated forms:a nonphosphorylated form (P0) and phosphorylated forms (P1, P2, and P3). Incubation of VSMCs with a high glucose level significantly increased the density ratio of P3/P0 as compared to a normal glucose level. Similarly, treatment of the cells with TPA significantly increased the P3/P0 ratio. Then, the increase in P3/P0 density ratio by a high glucose level was restored to the normal range by both staurosporine and calphostin C. These results suggest that high glucose levels may induce the reduction of GJIC activity in cultured VSMCs through excessive phosphorylation of Cx43 mediated by PKC activation. This may contribute to the development of diabetic angiopathy.
Next, we examined the phosphorylation state of Cx43 in the heart of streptozotocin-induced diabetics. The P3/P0 ratio of rat heart was significantly increased in the diabetic rats. To determine whether electrical conduction was affected by excessive phosphorylation of Cx43 in the diabetic rat heart, electrocardiogram was performed. QRS duration was significantly longer in the diabetic rats than their controls. Then, the administration of a PKCβ inhibitor, LY333531, to the diabetic rats completely normalized the prolonged QRS duration. The impaired electrical conduction by decreased gap junction activity may contribute to the pathogenesis of arrythmia and contractile dysfunction shown in diabetics.

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背景: 長期腹膜透析患者では, 腹膜表面を覆う中皮細胞の脱落と細動・静脈病変, 間質の線維性肥厚を中心とする組織学的変性を認め, 腹膜透析の除水不全の一因となっている. 腎性貧血患者に投与するerythropoietin (Epo) は, 造血作用以外にも血管新生作用を有することが報告されている. 今回, クロールヘキシジングルコネート (CG) を用いた腹膜硬化モデルラットにEpoを併用することで, 組織侵襲期におけるEpoの血管新生の組織への影響について検討を行った. 方法: 24匹の8週齢のSprague-Dawleyラットを4群に分け, (1) 腹腔内にCGとEpoの投与を行ったCG+Epo群, (2) CG投与のみを行ったCG群, (3) Epo投与のみを行ったEpo群と (4) 対照群を作成した. 投与開始28日後に屠殺し, 採取した壁側腹膜を用い, hypoxia-inducible factor-1α (HIF-1α), erythropoietin receptor (EpoR), pimonidazoleの発現を免疫組織学的に検索した. HIF-1α, vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), connective tissue growth factor (CTGF) のmRNA発現をReal-time PCRで評価した. またCD34とα-smooth muscle actin (α-SMA) の蛍光二重染色を行い, 血管密度・成熟度を計測した. 結果: CG群において, 腹膜の線維性肥厚が認められ, Epoの併用で抑制された. CG+Epo群のヘマトクリットはCG群に比べ著名に上昇した. CG群の肥厚した腹膜内にHIF-1α, EpoR, pimonidazole陽性細胞が多数存在し, Epoの併用により減少した. CG群では, CG+Epo群に比べHIF-1α・VEGF・Ang-2・CTGFmRNA発現の亢進を認め, CG+Epo群と比べると, これらのmRNA発現の有意な抑制を認めた. CG群のAng-1mRNAの発現は, CG+Epo群に比べると有意に抑制されていた. CG+Epo群の血管密度と血管成熟度は, CGのそれらに比べ有意な上昇を示した. 結語: CGの投与を行った腹膜硬化モデルラットにEpoを併用することで腹膜の線維性肥厚は, 抑制された. その理由として, Epoの造血作用と成熟血管網の構築を高めることで, 虚血組織への酸素供給量が上昇したことが挙げられる. また, Epo投与は低酸素状態を改善し, HIF-1α発現抑制を介したCTGF発現を抑えたと思われた.

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乳房温存オンコプラスティックサージャリーWGでは, oncoplastic breast-conserving surgery (以下OPBCS) に関する手技習得のためのステップアップガイドを作成中である。本稿ではそのなかのVolume displacementステップ1について解説する。

通常の乳房部分切除術においても, 皮下および大胸筋前面の剥離によって乳腺弁または脂肪乳腺弁を移動して欠損部を充填する方法は行われるが, ステップ1はさらに皮膚切開部位の選択や工夫, 乳頭乳輪複合体 (nipple-areolar complex, 以下NAC) の位置またはサイズを調整することで, 整容性の向上やNACの偏位・変形を防ぐための工夫を加えた手技である。

NACの偏位・変形を予防するための一つの方法として, 乳輪周囲の脱上皮化を併用して, 部分切除と反対方向にNACを移動することがある。脱上皮化を利用したNACの移動は, OPBCSの入り口ともいえる基本的かつ大切な手技なので, 是非とも習得していただきたい。

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本邦に適したoncoplastic breast-conserving surgery (OPBCS) に関する手技習得ステップアップガイドの最初のステップであるBasic OPBCSを解説する。

Basic OPBCSは, OPBCSに臨むにあたっての基本的な心構えである。

OPBCSでは, 腫瘍の大きさ, 広がり, 位置, 乳房の大きさ, 腫瘍/乳房容積比を評価することに加え, 以下の項目も考慮することが望まれる。

(1)背景の乳房構成 (脂肪性の乳房, 高濃度乳房) と皮膚の伸展性の評価

(2)立位での乳房の全体的形態や下垂の有無とその程度の評価

(3)乳頭・乳輪サイズと位置の評価や乳頭, 乳房下溝線および皮膚切開線等のマーキング

(4)術前 (立位) , 術中, 術後 (立位) の写真撮影による評価

(5)切除検体の重量 (容量) 測定

(6)術前ケア (喫煙, 糖尿病等の併存疾患の有無確認と管理) , 術後ケア (下着や保湿指導など)

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歯周疾患のリスクファクターとして糖尿病が注目されており,日本人における糖尿病患者の約95%はII型糖尿病に属していることから,II型糖尿病と歯周疾患との関連性を明らかにすることが求められている.高血糖状態で形成が促進されるadvanced glycation end products(AGEs)は,細胞表面受容体を介して認識され細胞内酸化ストレスの産生を亢進させ,種々のサイトカインや増殖因子の分泌を促進し,炎症反応を惹起する.しかし,AGEsが歯周組織の早期創傷治癒過程に及ぼす影響に関する報告はほとんどない.創傷治癒にかかわる主要因子のIII型コラーゲンは創傷治癒の早期に毛細血管や細胞の足場となり,組織の成熟化に深く関与している.そこで本研究では,歯周組織の創傷治癒過程に糖尿病が及ぼす影響を明らかにするために,II型糖尿病モデルのGoto-Kakizaki(GK)ラットの臼歯歯周組織に作製した人工的欠損の治癒過程におけるIII型コラーゲンおよびAGEsの局在を免疫組織化学的に観察した.生後8週齢のSD系ラットを対照群,GKラットを実験群とし,両群ともに歯肉全層弁を作製し,直径1mmのラウンドバーを用いて骨-セメント質欠損を作製した.人工的歯周組織欠損作製後,歯肉弁を元の位置に戻し1糸縫合を行い,術後3,5,7,14日にラット各5匹を安楽死させ,10%中性緩衝ホルマリンにて灌流固定後,被験歯根を含む周囲組織を一塊として採取し脱灰した.脱灰後,パラフィン切片を作製し,III型コラーゲンおよびAGEsの免疫染色,H-E染色を行い,光学顕微鏡で観察した.その結果,術後5日および7日においては,対照群に比べて実験群では歯周組織欠損部に形成された肉芽組織におけるIII型コラーゲンの発現は少なかった.術後14日において,対照群に比べて実験群ではIII型コラーゲンが不規則に配列されていた.また,すべての実験期間を通して実験群では,類円形を呈さない不定形な管腔を形成する毛細血管が増生しており,その血管内皮細胞にはAGEsの局在が認められた.以上の結果から,AGEsは血管内皮細胞上に存在する受容体(receptor for AGEs:RAGE)によって認識された後,細胞内酸化ストレスの産生を亢進して線維芽細胞および毛細血管構成細胞に作用し,III型コラーゲンの産生を低下させ,毛細血管形成不良も惹起することで,歯周組織欠損部における早期創傷治癒過程に悪影響を与えていることが示唆された.

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The goals of the study were to investigate the effects on bone bioactivity of a titanium dioxide layer formed by hydrothermal oxidation of a titanium surface with hydrogen peroxide（H₂O₂） and loading with fibroblast growth factor-2（FGF-2） in vitro and in vivo.
　Ti-6Al-4V discs were hydrothermally oxidized with H₂O₂ and then loaded with FGF-2. After cytotoxicity testing, Ti-6Al-4V mini-implants were subjected to the same treatment, and their osteogenic potential was evaluated histologically in a rat model.
　H₂O₂ hydrothermal oxidation resulted in a dense porous network structure and hydrophilic changes, which improved retention of FGF-2. Morphologically, the cell density was higher, cell elongation was more pronounced, and the cell adhesion area was significantly higher in FGF-2-loaded cells than in those without FGF-2. In a cell proliferation assay using mouse osteoblast-like cells, absorbance tended to increase over time, especially in the FGF-2 group after 7 and 14 days. In a bone differentiation assay based on ALP activity, there was a significant increase in the FGF-2 group after 14 days. In the rat model, H₂O₂ hydrothermal oxidation and FGF-2 loading both resulted in more laminar bone tissue in the bone marrow around the mini-implant.
　These results suggest that titanium surface functionalization using H₂O₂ hydrothermal oxidation and FGF-2 may promote initial cell adhesion, proliferation and osteodifferentiation, and enhance bone bioactivity. These effects all contribute to the early bonding of an implant with the surrounding bone.

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Adenosine 3′, 5′-cyclic monophosphate (cyclic AMP), discovered by Sutherland et al. in 1958, is now considered to be the secondary messenger of various β-adrenergic actions, such as cardiac inotropic action, glycogenolysis, lipolysis and relaxation of several smooth muscles. (1) Many investigators have already detected the presence of inhibitory β-adrenergic receptors in the vas deferens of some mammals. (2) Phosphodiesterase inhibitors (i. e. aminophylline, theophylline and papaverine) can reduce or abolish contractions of the vas deferens or rabbits, guineapigs and rats. (3) Externally applied dibutyryl cyclic AMP, a derivative of cyclic AMP, can reduce or abolish contractions of the vas deferens of guinea-pigs, rats and humans.
From the above-mentioned previous reports by many authors, it may be suggested that cyclic AMP is the intracellular mediator of inhibitory β-adrenergic action of the vas deferens smooth muscle.
In this paper, changes in tissue cyclic AMP levels of the rat vas deferens following administration of catecholamines, adrenergic blocking agents and aminophylline have been observed.
The methods were as follows:
1) 9 weeks old male Wistar rats (216±21g), anesthetized with sodium pentobarbital 50mg/kg i. p., underwent lapalotomy.
2) A drug or drugs were injected into the vena cava inferior.
3) 3, 8, 15, 30 or 60 minutes after the injection, the total length of both vas deferens was excised.
4) Immediately cyclic AMP levels of each tissue were estimated, using the cyclic AMP Assay Kit (Radiochemical Centre, Amersham, England).
The saline-injected group and drug-administered group were compared statistically.
The results obtained were as follows:
1) 20μg/kg of epinephrine or isoproterenol could elevate tissue cyclic AMP levels maximamly 8 minutes after the injection of these drugs.
2) The elevation of tissue cyclic AMP levels induced by the injection of epinephrine or isoproterenol was dose-dependent.
3) Preinjection of propranolol 1.0mg/kg abolished the elevation of tissue cyclic AMP levels induced by the injection of epinephrine 20μg/kg or isoproterenol 20μg/kg. But, preinjection of phenoxybenzamine 1.0mg/kg caused no effects.
4) Aminophylline 50mg/kg neither changed the basal tissue cyclic AMP level nor acted synergistically when combined with epinephrine or isoproterenol.
5) Propranolol 1.0mg/kg, phenoxybenzamine 1.0mg/kg or norepinephrine 0.2-200μg/kg did not change the basal tissue cyclic AMP level.
My results may add some proof to the hypothesis that cyclic AMP is the intracellular mediator of inhibitory β-adrenergic action of vas deferens smooth muscle.

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Electrophysiological studies were made on blinking in unanesthetized rabbits as to its development mechanism.
1. "Blink r eflex" was observed to developed unilaterally on the eye after a certain amount of air pressure was given to that eye's cornea and recorded by electromyography.
2. The frequency of developing "blink reflex" was either increased or decreased following the wearing of a contact lens.
3. In EEG there developed arousal reaction following a 100 c/s stimulation on reticular formation (RF), nucleus centrum medianum (CM) and nucleus ventralis anterior (VA), recruiting response following an 8 c/s stimulation on CM and VA, and spike and wave following a 3 c/s stimulation on CM and VA. Such development was always accompanied with bilateral blinking.
4. The freq u ency of blinking developing due to central nerve stimuli either increased or decreased following the wearing of a contact lens.
5. Three types of blinking developed following c entral nerve stimuli, T type which was continuous for a certain duration at certain intervals in the post -stimulation stage, P type which was repetitious during and immediately after stimulation, and TP type which was a mixed form. Generally T type often developed in association with spike and wave, P and TP type tended to develop in association with recruiting response and arousal reaction.
6. Th e types of evoked muscular discharge following central nerve stimuli maintained almost a parallel relationship with the types of blinking.
7. A comparative study of the effects of barbiturate, myanesin and chlorpromazine on "blink reflex" and flexion reflex revealed that both reflexes were of the same origin of mechanism.
8. Experimentally barbiturate, myanesin and chlorpromazine worked differently on blinking due to central nerve stimuli and "blink reflex". It was assumed that these two kind of blink were of different origin.
9. The inhibitory action of barbitura te, myanesin and chlorpromazine on blinking due to central nerve stimuli showed a parallel relationship with their inhibitory acti on on uusal reaction, recruiting response and evoked muscular discharge.

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