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We previously developed a new simple technique of methylene blue (MB) staining for the discrimination of aluminum (Al)-tolerant protoplasts from 4 plant species (rice, oats, maize and pea). The objectives of the present study were to confirm the applicability of this technique to a wider range of plant species, cultivars and lines, and to identify a common strategy for the early stage of Al tolerance. A total of 10 plant species, two Brachiaria spp., two Oryza spp., buckwheat, maize, pea, triticale, wheat and barley, corresponding to 18 different plant samples (species, cultivars, and lines), were used. Al tolerance (relative net root elongation of the longest root), which was screened at 20 μM AICI3 in 0.2 mM CaCl2 (pH 4.9) for 24 h, ranged widely from 10 to 88. Among cultivars and lines within the same species, Al accumulation in the root tip portion was higher in Al-sensitive plants, corresponding to more severe permeabilization of the plasma membranes (PM). Protoplasts isolated mainly from the epiderm...

Objective. The aim of this study was to evaluated the outcomes of living related kidney transplantation in small children. Materials and methods. Ten pediatric patients with body weights less than 10 kg received parental kidney transplants (five mothers and five fathers). An intra-abdominal approach was used in nine children and a retroperitoneal approach in one child. Bilateral, left, or right nephrectomy was performed in seven, two, and one child, respectively. Immunosuppression consisted of either cyclosporine (n = 7) or tacrolimus (n = 3) with either mizoribine (n = 4) or mycophenolate mofetil (MMF) (n = 5) or azathioprine (n = 1), and methylprednisolone (n = 10). Antilymphocyte globulin was used in the first series of four children; basiliximab in the most recent five children. Results. All renal allografts functioned immediately after transplantation despite the mismatched size of the large renal allografts. Nine of 10 children were alive with a functional allograft at 6 to 196 months posttransplantation. One child died of intra-abdominal bleeding 5 days posttransplantation. One child has suffered chronic allograft nephropathy 11 years posttransplantation (serum creatinine 3.3 mg/dL). The remaining eight children display good renal function (serum creatinine = 0.2 to 1.43 mg/dL). Steroids were withdrawn in eight of nine children; one child continues on alternative-day therapy. One child (LD55) exceeded the mean standard height. The most recent height standard deviation (SD) scores were superior (-1.75 ± 1.39 [-3.83 to 0.54]; P

The order of aluminum (Al) tolerance in triticale lines (ST2>ST22) after re-elongation in an Al-free 0.2 mM calcium (Ca) solution for 9 h (Ca period) following 1 h pretreatment with 20 μM Al (Al period) agreed with that after 24 h of Al treatment. Permeability of the plasma membrane (PM) of root-tip cells after the Ca period was significantly increased in Al-sensitive ST22. Al was accumulated more heavily in the root-tip portion of ST22 than in that of ST2, although similar amounts of malic and citric acid anions were released from both triticale lines. We established a new system examining lipid permeability using synthesized nylon-2,8 ultrathin and porous capsules trapped previously with 0.1% (w/v) methylene blue solution and coated thereafter with PM lipid isolated from root tips by a newly developed technique. Permeability of the PM lipid measured with time in 0.2 mM Ca with or without 50 μM Al photometrically (A 680) was significantly greater in Al-sensitive ST22 after 5 min of Al treatment. This is ...

Aims Approximately 80% of uracil is excreted as β-alanine, ammonia and CO2 via three sequential reactions. The activity of the first enzyme in this scheme, dihydropyrimidine dehydrogenase (DPD), is reported to be the key determinant of the cytotoxicity and side-effects of 5-fluorouracil. The aim of the present study was to re-evaluate the pharmacokinetics of uracil and its metabolites using a sensitive assay and based on a newly developed, physiologically based pharmacokinetic (PBPK) model. Methods [2–13C]Uracil was orally administrated to 12 healthy males at escalating doses of 50, 100 and 200 mg, and the concentrations of [2–13C]uracil, [2–13C]5,6-dihydrouracil and β-ureidopropionic acid (ureido-13C) in plasma and urine and 13CO2 in breath were measured by liquid chromatography–tandem mass spectrometry and gas chromatograph–isotope ratio mass spectrometry, respectively. Results The pharmacokinetics of [2–13C]uracil were nonlinear. The elimination half-life of [2–13C]5,6-dihydrouracil was 0.9–1.4 h, whereas that of [2–13C]uracil was 0.2–0.3 h. The AUC of [2–13C]5,6-dihydrouracil was 1.9–3.1 times greater than that of [2–13C]uracil, whereas that of ureido-13C was 0.13–0.23 times smaller. The pharamacokinetics of 13CO2 in expired air were linear and the recovery of 13CO2 was approximately 80% of the dose. The renal clearance of [2–13C]uracil was negligible. Conclusion A PBPK model to describe 13CO2 exhalation after orally administered [2–13C]uracil was successfully developed. Using [2–13C]uracil as a probe, this model could be useful in identifying DPD-deficient patients at risk of 5-fuorouracil toxicity.

We reviewed the transplantation data and results of histopathological studies with additional angiotensin II (AII) immunostaining of renal graft biopsies of nine cases (10 grafts) with recurrent proteinuria and three controls without recurrent proteinuria that received renal transplantation for primary focal segmental glomerulosclerosis (FSGS) between 1986 and 2002. Recurrent FSGS was confirmed in six grafts from nine cases by light microscopy. In cases with recurrent proteinuria, loss of graft function was noted in all six renal grafts received between 1986 and early 1992 but in none of four grafts received between late 1992 and 2002. Two of four patients of the late group but none of those of the early group received angiotensin converting enzyme (ACEI) or angiotensin II receptor blocker (ARB) with plasma exchange (PE). In control cases without proteinuria, AII immunostaining was detected in tubules but not in glomeruli in 1-hour biopsies as well as later on. In cases with recurrent proteinuria, AII immunostaining was detected in both tubules and glomeruli, although glomerular AII staining was not observed in 1-hour biopsies. Our results suggest that effective treatment of post-transplantation recurrent FSGS requires ACEI or ARB with PE in the absence of another etiology.

Procarboxypeptidase R (proCPR), also known as thrombin-activatable fibrinolysis inhibitor (TAFI), is present in plasma and can be activated to carboxypeptidase R (CPR) by trypsin-like enzymes such as thrombin and plasmin. CPR has the carboxypeptidase B-like activity that can inactivate the inflammatory peptides such as C5a by removing the C-terminal arginine and can interfere with fibrinolysis by removing C-terminal lysine residue of fibrin. In the present study, we conducted to produce monoclonal antibodies (mAbs) by using spleen cells from proCPR-deficient mice immunized by partially purified mouse proCPR. The mAbs obtained were IgM isotype and reacted with proCPR and interfered with activation of proCPR to CPR by thrombin-thrombomodulin complex. Some BALB/c mice implanted with the hybridoma died in 7 days, and intravenous injection of the mAb to BALB/c mice induced transient elevation of GOT and GPT in plasma although injection to the deficient mice did not. Furthermore, the histological features showed the focally lesions in liver tissue of BALB/c mice injected with the mAb. Since liver is the major site of proCPR synthesis, IgM mAb to proCPR should have induced local inflammation at the side resulting in induction of hepatitis.

It has been suggested that plasma membrane (PM) lipids play a major role in aluminum (Al) tolerance; however, no direct investigations have been carried out using PM lipids from root-tips. Here we report a new technique for PM isolation as an alternative to the laborious two-polymer phase partitioning method that is commonly applied, as follows: 1) separation of protoplasts from 1-cm root-tip portions by enzymatic digestion, 2) attachment of the purified protoplasts to glass plates coated with polylysine, 3) preparation of PM ghosts by successive burst of the attached protoplasts using three separate buffer solutions (25 mM PIPES, 5 mM EDTA, and 2 mM MgCl2, at pH 7.0) with slow stirring for 60 s. The PMs were confirmed to be devoid of organelle membranes by fluorescence microscopy, thin layer chromatography (TLC) and western blot analysis. The PM lipids obtained were found to be useful for studies on their differential permeability and lipid composition between lines of triticale or cultivars of maize und...

子宮内膜癌の消化管転移は比較的稀であり,なかでも大腸転移の本邦報告例は極めて少ない.今回われわれは,子宮内膜癌術後に直腸転移をきたした1例を経験したので報告する.症例は77歳,女性,平成12年4月に子宮内膜癌の診断で子宮全摘両付属器切除術を施行した.外来フォロー中, 2年7カ月後に近医で右胸部異常陰影,便潜血陽性を指摘され当院を紹介された.精査の結果,原発性直腸癌と診断し,胸部異常陰影は直腸癌の肺転移を疑ったが確診には至らず, 3カ月経過観察した後に直腸切断術を施行した.術後病理所見にて直腸の病変は主座が粘膜下層から固有筋層に存在し,組織像が平成12年時の子宮内膜癌の所見に酷似しており,子宮内膜癌の直腸転移と診断した.術後外来にてMPA (酢酸メドロキシプロゲステロン)内服で経過観察中であったが,直腸術後11カ月で他病死した.

We report a case of prostatic cystadenoma. A 81-year-old man visited our hospital with the chief complaint of urinary difficulty. RUG and DIP showed bladder and prostatic urethra deviated to the left side. TRUS, CT and MRI revealed multilocular cyst in the pelvic cavity. Resection of the cystic wall was performed under the diagnosis of pelvic cyst. The cysts were originated from the right side of the prostate and adhered with the bladder severely. The histopathological diagnosis was prostatic cystadenoma. Fortyfive cases of prostatic cyst in the Japanese literature are reviewed.

症例は59歳,男性.腹痛を主訴として来院. WBC 26,600/μl, CRP 2,72mg/dlと上昇,腹部CTでは骨盤入口部に壁の一部にdensityの上昇を伴う腫瘤が認めらた.開腹所見では回腸未端から約50cm口側に,腫大した回腸と隣接した腸間膜に膿瘍を認めたため,同部位を切除した.切除された回腸に,膿瘍と交通する瘻孔を認めた.病理組織所見では,瘻孔周囲に腸粘膜に相当する組織がみられたため,回腸憩室の腸間膜への穿通による膿瘍形成と診断された. Meckel憩室を除く小腸憩室は剖検で0.006～1.3%と非常に稀である.消化管穿孔による膿瘍形成のCT所見の特徴は,腸管に隣接した内部にガス像を含む比較的high densityの嚢胞腔と報告されているが,本症例でも同様の所見が得られた.

The human genome has been sequenced, and investigation of its products has become possible in a sequence-based framework. More than 200,000 protein species are expressed in the body from approximately 30000 human genes. The term proteome, coined as a linguistic equivalent to the concept of genome, is used to describe the complete set of proteins that is expressed, and modified following expression, by the entire genome in a cell at any one time. Protein types and amounts expressed in a body vary greatly depending upon whether it is healthy or ill. Therefore, proteomics is attracting an increasing interest in its application to better understanding of disease processes, to development of new biomarkers for diagnosis and early detection of disease, and to accelerate drug development. There are numerous opportunities for medicine, although it is quite challenging to meet the needs for high sensitivity and high throughput required for disease-related investigations.

Background and Aims Sperm motility is regulated by protein phospborylation. The 66 kDa protein obtained from hamster sperm flagella was phosphorylated at serine residues associated with the motility initiation. In order to understand the regulatory mechanism of sperm motility, the 66 kDa protein was identified in the present study.

A 43-year-old woman with end-stage renal disease originating from IgA nephropathy entered chronic haemodialysis therapy. She then received an ABO-incompatible living related renal transplantation. Initial immunosuppression consisted of azathioprine, methylprednisolone and tacrolimus. At 155 days after transplantation, the azathioprine was changed to mycophenolate mofetil for continuous graft dysfunction. Furthermore, a total of three courses of anti-rejection therapy was given. At 665 days after transplantation, diagnosis of BK-virus nephropathy was made by immunohistochemical analysis and viral DNA assay. Therefore the immunosuppression therapy was reduced for graft dysfunction. All five renal biopsy specimens were examined retrospectively in order to determine when the BK virus nephropathy had developed. The expressions of SV40 large T antigens were detected from the third (117 days) to the fifth (665 days) biopsies, with increasing numbers of SV40 large T antigen positive cells. In addition, many cells contained inclusion bodies which were already present in the urinary sediment for 3 months post-transplantation. Although it is difficult to make a diagnosis of early stage of BKVN, we have to consider with caution if urinary cells with inclusion body are seen. Awareness of BKVN at the earliest opportunity is important in order to avoid over-immunosuppression.

In clinical and diagnostic proteomics, it is essential to develop a comprehensive and robust system for proteome analysis. Although multidimensional liquid chromatography/tandem mass spectrometry (LC/MS/MS) systems have been recently developed as powerful tools especially for identification of protein complexes, these systems still some drawbacks in their application to clinical research that requires an analysis of a large number of human samples. Therefore, in this study, we have constructed a technically simple and high throughput protein profiling system comprising a two-dimensional (2D)-LC/MS/MS system which integrates both a strong cation exchange (SCX) chromatography and a microLC/MS/MS system with micro-flowing reversed-phase chromatography. Using the microLC/MS/MS system as the second dimensional chromatography, SCX separation has been optimized as an off-line first dimensional peptide fractionation. To evaluate the performance of the constructed 2D-LC/MS/MS system, the results of detection and identification of proteins were compared using digests mixtures of 6 authentic proteins with those obtained using one-dimensional microLC/MS/MS system. The number of peptide fragments detected and the coverage of protein sequence were found to be more than double through the use of our newly built 2D-LC/MS/MS system. Furthermore, this multidimensional protein profiling system has been applied to plasma proteome in order to examine its feasibility for clinical proteomics. The experimental results revealed the identification of 174 proteins from one serum sample depleted HSA and IgG which corresponds to only 1 microL of plasma, and the total analysis run time was less than half a day, indicating a fairly high possibility of practicing clinical proteomics in a high throughput manner.

Aims: The major transmission route of Helicobacter pylori remains unclear. In this study, we examined H. pylori in the environmental waters in Japan. Methods and Results: A total of 24 water samples were collected from the upper, middle and downstream reaches of four Japanese rivers. Helicobacter pylori-specific DNA was examined using nested PCR. In addition, 224 children who lived near one river were studied by the stool antigen test for H. pylori prevalence. Helicobacter pylori DNA was detected in the water from the middle and downstream reaches of all four rivers, but not in the upper reaches. Helicobacter pylori was not found in cultured water samples with positive PCR results. Helicobacter pylori prevalence in the children examined was 9·8% for those living near the middle reaches and 23·8% nearby downstream, both of which were higher than the value in an area distant from the river (0%) (both, P

In our previous studies (Fujinoki et al., 2001, 2003), we reported that two types of 36 kDa proteins, designated 36K-A protein and 36K-B protein, obtained from hamster sperm flagella, are associated with motility activation and phosphorylated in a cAMP-dependent manner at serine residues. In the present experiments, we focused on the hamster (Mesocricetus auratus) 36K-A protein, which was analyzed by peptide mass finger printing and amino acid sequencing. The results suggest that 36K-A protein is a pyruvate dehydrogenase E1 component beta subunit lacking the N-terminal 30 amino acids. Moreover, our results suggest that 36 K-A protein is localized in the fibrous sheath of the principal piece of hamster spermatazoa.

症例は83歳,男性.喘息発作と腹痛を主訴に来院.腹部は著明に膨満し下腹部を中心に腹膜刺激症状を認めた.腹部単純X線で腹腔内遊離ガスを認めたため,腸管穿孔による腹膜炎の診断で手術を施行した.手術所見では横隔膜,小腸壁に広範な気腫性変化をきたしており一部穿孔を疑う部位を含め小腸部分切除を施行した.病理所見では粘膜下層から筋層,また漿膜下層に嚢胞が存在し,嚢胞周囲に線維性の肥厚,肉芽性の変化を認めた.以上より小腸の嚢腫様気腫症と診断された.本症の治療は高圧酸素療法などの保存的治療が中心であるが,腹膜刺激症状を有する場合は消化管穿孔との鑑別が重要と考える.

症例は46歳男性, 胸痛を自覚し受診.胸部X-pにて右血気胸と診断, 胸腔ドレーンを挿入したところ, 1200mlの暗血性胸水の流出を認めた.胸部X-p, 胸部CTにて右上葉に結節影を認めた.血気胸に対し胸腔鏡下手術を行った.壁側胸膜に古い出血点を確認.肺尖部のブラの直下に径1.5cmの結節性の腫瘍を認め, 腫瘍も含む右肺尖部分切除術を施行した.術後病理組織検査にて大細胞癌の診断となり, 初回手術2週後に開胸にて右上葉切除術を施行した.術後Vindesine (VDS), Cisplatin (CDDP) による化学療法を2クール施行.術後1年10ヵ月経過した現在再発徴候なく健在である.血気胸を契機に原発性肺癌が発見される例はまれである.肺癌に合併する気胸に関して若干の文献的考察を加えて報告する.

症例は62歳女性.近医より胃粘膜下腫瘍(SMT)精査目的で当院紹介入院.内視鏡所見では,胃噴門直下後壁に径約2 cm大のSMTを認め,中心陥凹などの悪性を示唆する所見なく, EUS・CT像でも内部均一な腫瘤として描出された.術前診断としては平滑筋腫などの良性筋原性腫瘍もしくは低悪性度GISTなどを考え,本人・家族と相談の上,消化管内視鏡下切除を行った.切除標本の病理組織所見では,紡錘形細胞の錯走像からなり,核分裂像は1/50HPF,免疫染色にてc-kit・CD34ともに陽性を示す狭義のGISTでborderline malignancyと診断された.胃GISTを消化管内視鏡下に切除した症例を経験したため,その診断および治療の過程を報告し,当院における胃SMTに対する治療方針についても言及した.

We discuss a patient with Alport syndrome who received a renal transplant from a donor with thin basement membrane disease. A 30-year-old woman, diagnosed with Alport syndrome on the basis of sensorineural hearing loss, characteristic renal biopsy findings and a family history of microhaematuria, entered chronic haemodialysis therapy. She then received a renal transplant donated from her father, who had sensorineural hearing loss and persistent microhaematuria. On the day of renal transplantation, a 1-h graft biopsy after reperfusion showed thin basement membrane disease. We re-tested the patient's native kidney biopsy specimen by immunohistochemical staining using alpha-chain-specific collagen type IV monoclonal antibodies. There was no expression of collagen type IV alpha3-, alpha4- and alpha5-chain on glomerular basement membrane, but positive staining of alpha5-chain on Bowman's capsular basement membrane was noted. A diagnosis of autosomal-recessive Alport syndrome was made. We concluded that this family might display different phenotypic expressions of the same genotype: one suffered end-stage renal disease and the other thin basement membrane disease.

In our previous paper [M. Fujinoki et al. (2001) BIOMED: Res. 22, 45-58], we reported that two types of 36-kDa protein, which were designated as 36K-A protein and 36K-B protein, obtained from hamster sperm flagella were phosphorylated at serine residues associated with the regulation of motility activation. In the present experiments, it was suggested that these two types of 36-kDa protein were phosphorylated in a cAMP-dependent manner associated with motility activation of hamster spermatozoa. Because the 36K-B protein was the most intensely phosphorylated in a cAMP-dependent manner, attempts were made to further characterize it. The 36K-B protein was assumed to be localized in the middle piece. The localization of the 36K-B protein was the same as that of the 36-kDa protein reported in our previous paper [Y. Si et al. (1999) Mol. Reprod. Dev. 52, 328-334]. In order to identify the 36K-B protein, it was analyzed by peptide mass finger printing and amino acid sequencing. The results suggested that the 36K-B protein was a pyruvate dehydrogenase E1 component beta subunit and a component of the mitochondrial sheath of the middle piece.

Human IgM antibody (Ab) to gangliosides induced cytolysis of HIV-1-infected cells by homologous human complement. We expected that any human IgM Ab reactive with HIV-1 infected cells could cause complement-mediated cytolysis. The trans-chromosome mouse (TC mouse) contains human chromosomes harboring genes responsible for immunoglobulin production. Spleen cells from TC mice immunized with recombinant Nef were fused with mouse myeloma cells to generate hybridomas, and we selected those that produced human mu-chain-positive Abs reactive with Nef fixed on an ELISA plate. However, the L-chain of the monoclonal Abs (mAbs) were murine lambda in type and were chimeric, and we could not succeed in obtaining mAb with human mu- and human kappa-chains. The chimeric mAbs reacted with the HIV-1 infected cells as seen with flow cytometric analysis, and the surface expression of Nef was also detectable on chronically infected OM10.1 cells which had no detectable gp120. However, although the reaction of the chimeric IgM mAb with HIV-1-infected MOLT4 cells induced C3 deposition on cell surfaces on incubation with fresh human serum, the cells remained unlysed, as determined by 51Cr release assay. The amount of Nef antigen on the cells might not have been high enough to overcome the function of HRF20 (CD59) that restricts formation of membrane attack complexes of homologous complement. However, combination of anti-Nef IgM mAb with other IgM mAbs reactive with the surface of HIV-1-infected cells may induce a synergistic effect in complement mediated cytolysis.

進行乳癌に対するpaclitaxel治療中に間質性肺炎を発症し死亡した1例を経験した.症例は61歳の女性.呼吸苦を主訴に当院呼吸器内科を受診したが,皮膚浸潤を伴う乳腺腫瘍と両側性の胸水貯留を指摘され当科へ紹介された.癌性胸膜炎を伴う進行乳癌と診断し175mg/m2/月のpaclitaxel治療を開始した.第1クール終了後にgrade 3の白血球減少と好中球減少を認めたが他には特に副作用を疑わせる所見は認めなかった.第2クール終了時には胸水も著明に減少し呼吸苦も消失したが,数日後に両側性の間質性肺炎を発症した.ステロイド投与を開始したが改善することなく4日後に死亡した.他に原因となる要素がないことなどから, paclitaxelの副作用によるものと考えられた.乳癌に対するpaclitaxel治療で発症した間質性肺炎の報告は極めて少ないが,生じうる副作用として留意する必要があると考えられた.

症例は76歳,男性. 2000年11月,眩暈,ふらつきを主訴に受診.血液検査にてHb 6.8 g/dlと著明な貧血を認めた.下部消化管内視鏡検査では上行結腸に単発の大きさが約15 mm大の鮮紅色を伴った円形の隆起性病変を認めた.腹部血管造影では,回結腸動脈から上行結腸中部を栄養する分枝に斑状の強い濃染像を認めた.他部位に貧血をきたす病変はなく,上行結腸angiodysplasiaの出血とそれに伴う高度の鉄欠乏性貧血と診断し,腹腔鏡補助下上行結腸切除術を施行した.術後経過は良好で術後23カ月の現在,出血,貧血の兆候は認めていない.

We evaluate clinical characteristics of primary vesicoureteral reflux in infants in a multicenter study in Japan with special reference to the relation of renal parenchymal damage to urinary tract infection and gender.Infants younger than 1 year old with primary vesicoureteral reflux were recruited from 14 hospitals during the 3-year registration period beginning in January 1996 and ending in December 1998. Various clinical parameters as well as renal parenchymal lesion on dimercaptosuccinic acid scintigraphy were evaluated.Of 356 infants enrolled 296 (83%) were male and 60 (17%) were female. In 85% of infants presenting symptom was febrile urinary tract infection. There were 204 bilateral (57%) and 152 unilateral (43%) cases. Reflux was bilateral in 56% of males versus 65% of females, and high grade (grades IV and V) in 58% of males versus 55% of females. Diffuse parenchymal lesion was similarly noted in infants with or without prior urinary tract infection (38% and 46%, respectively) and was more often noted in male than in female infants (42% versus 25%).Despite the current use of screening prenatal ultrasound, many infants are still diagnosed as having vesicoureteral reflux only after the occurrence of urinary tract infection. The greater severity of renal parenchymal lesion in male infants combined with similar incidence of diffuse parenchymal lesion in those with or without prior infection suggests preexisting congenital abnormalities in the male refluxing kidney.

Cyclic AMP-dependent phosphorylation of tyrosine residues in a protein with a molecular mass of 15-kDa is thought to play a key role in the initiation of sperm motility in salmonid fishes (7). In this study, flagellar proteins were solubilized with 7 M urea and fractionated by sucrose density-gradient isoelectric focusing. A band relevant to 15-kDa was detected in the fraction at pH 5 on electrophoresis and analyzed using phosphotyrosine antibody. Analysis of the band by reversed phase chromatography showed a single peak, suggesting that the 15-kDa protein was completely purified. Amino acid sequence of His-Ile-Phe at the N-terminus was detected by analysis using peptide sequencer, and the fourth base could not be determined. Further analysis of the molecular structure of the protein by LC-MS/MS showed that partial amino acid sequence of the 15-kDa protein from tryptic fragments were similar to parts of the amino acid sequence of tubulin. The protein was not detected in immunoblots using anti-tubulin polyclonal antibody, suggesting that the 15-kDa protein may be not debris of the tubulin but a novel protein with a common partial sequence of tubulin.

Aims: To investigate the existence of Helicobacter pylori in cow's milk as one of the foods which most Japanese children eat. Methods and Results: Detection of H. pylori was demonstrated by the semi-nested polymerase chain reaction (PCR), a culture method and electron microscopy. Semi-nested PCR demonstrated the ureA gene of H. pylori in 13 of 18 (72·2%) raw milk samples and in 11 of 20 (55%) commercial pasteurized milk samples. Helicobacter pylori binding immunomagnetic beads with H. pylori-specific goat anti-H. pylori antibody was shown by electron microscopy in both raw and pasteurized milk positive for the ureA gene. Helicobacter pylori was cultured in one raw milk sample, whereas it was not cultured in pasteurized milk samples. Conclusions, Significance and Impact of the Study: There is a possibility that cow's milk is a transmission vehicle in childhood H. pylori infection, although we failed to confirm the survival of H. pylori in pasteurized milk.

The effect of composition on the dispersion of AuxCu1 − x bimetallic nanoparticles into a nylon 11 matrix has been investigated. TEM, UV–VIS, EDX, and XPS depth profiling were used for characterizing the changes in the composition of the bimetallic particles and the depth distribution of the particles in the nylon 11 layer caused by annealing in an N2 atmosphere. TEM observation and EDX analysis revealed that island-like bimetallic particles were formed on the nylon 11 surface before annealing, the composition of which was systematically controlled. Upon annealing in an N2 atmosphere, the AuxCu1 − x bimetallic particles with x ≤ 0.55 were not dispersed into the nylon 11 layer while those with x ≥ 0.70 were homogeneously dispersed in the films, indicating the existence of a critical composition for penetration of the bimetallic particles. By comparing the composition and structure of the bimetallic particles, the cause of these finding is discussed in terms of the surface free energy of the particles, which can be varied with their composition by the effect of alloying.

Carboxypeptidase R (EC 3.4.17.20; CPR) is an unstable basic carboxypeptidase found in fresh serum in addition to carboxypeptidase N (CPN) which is a stable enzyme. CPR in fresh serum is generated from its zymogen (proCPR) during coagulation by trypsin-like enzymes such as thrombin and thrombin/thrombomodulin complexes. Since removal of the C-terminal arginine abrogates the anaphylatoxin activity of C3a and C5a, CPR and CPN are regarded as anaphylatoxin inactivators. We report here that the culture supernatant of activated human neutrophils converts proCPR to CPR. Addition of an elastase specific inhibitor, N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl ketone (MSAAPVCK) to the supernatant of stimulated neutrophils completely inhibited activation of proCPR. On the other hand, a thrombin specific inhibitor, p-Nitrophenyl-p'-amidinophenyl-methanesulfonate hydrochloride (pNP-pAPMS) inhibited only 16% of proCPR activation by the neutrophil supernatant. Furthermore, purified elastase converted proCPR to CPR. Therefore, elastase can activate proCPR directly, or indirectly through activation of some proteases, which have been contaminating in reagents. Release of CPR generating enzymes from neutrophils should play an important role in regulation of excess inflammation.

We previously created a low-immunogenic core streptavidin mutant No. 314 (LISA-314) by replacing six amino-acid residues for use as a delivery tool for an antibody multistep pre-targeting process (Yumura et al., Protein Sci., 22, 213-221, 2013). Here, we performed high-resolution X-ray structural analyses of LISA-314 and wild-type streptavidin to investigate the effect of substitutions on the protein function and the three-dimensional structure. LISA-314 forms a tetramer in the same manner as wild-type streptavidin. The binding mode of d-biotin in LISA-314 is also completely identical to that in wild-type streptavidin, and conformational changes were observed mostly at the side chains of substituted sites. Any large conformational changes corresponding to the reduction of B factors around the substituted sites were not observed. These results demonstrated the LISA-314 acquired low immunogenicity without losing structural properties of original wild-type streptavidin.

Organizational patterns are known as the basis for Agile software development movement. Patterns usually form a network having relationships among them to support users understand and utilize patterns efficiently and effectively. However little is known about the nature of pattern networks, such as how are organizational patterns different from other patterns from the viewpoint of centrality. To clarify such characteristics, we mine a network consisting 285 patterns including 15 organizational patterns from an existing online pattern repository called Portland Pattern Repository. By applying network analysis techniques to the mined network, we revealed several interesting characteristics of the pattern network and organizational patterns such as that the degree centrality seems to somewhat reflect the commonness and generality of the corresponding pattern.

Recently, the inter-vehicle communication systems have been proposed for the prevention of car crash. By the way, a radiation pattern of car antenna is changed by effects of car body and antenna positions. Therefore, it is important to know the influence of car body and antenna positions on radio wave propagation characteristics. In the numerical simulation, from a viewpoint of the reduction of computational cost, it is interested to consider a simplified car model that can obtain reasonable numerical results. In this paper, effects of car body on radiation pattern of car antenna mounted on a side mirror for inter-vehicle communications are investigated. As a first step in this work, both strict and simplified models of car body are considered. As the numerical simulation method, the FDTD method is used.

Various urological complications in VATER association require careful management. A 15-year-old boy with VATER association, including a hypoplastic lower urinary tract and diphallia, presented with chronic kidney disease and incontinence after a right loop ureterostomy. In order to acquire urinary continence without renal function impairment, an ileocecal reservoir with umbilical catheterizable stoma was created as a urinary reconstruction. The ectopic posterior penis was resected for cosmetic reasons, and the stump of the hypoplastic urethra was opened at the perineal skin. Clean intermittent self-catheterization through the umbilicus using disabled bilateral limbs was then achieved. This report describes the management of VATER association in a patient with complicated urological anomalies.

Epigenetic regulations have been known as phenomena such as the X chromosome inactivation since the mid-twentieth century. Recently, DNA methylation and post-translational modification on histones, which consist chromatin, have been proved to mediate the epigenetic regulations. Then, DNA methyltransferases and histone deacetylases have been implicated in carcinogenesis to be attractive drug targets.

We examined embryonic carcinoma (EC) cells for a potential prototype molecule of C3, the third component of complement. PCR primers, corresponding to the base sequence derived from the C3 cDNA of several species, were used for PCR amplification of the EC cell cDNA. All the PCR products obtained had the same sequence and showed no sequence homology to C3. Subsequently, cDNA clones were isolated from a mouse liver cDNA library using the PCR product as a probe. Unexpectedly, neither the base sequence of the cDNA clones nor the amino acid sequence deduced from the cDNA showed homology to C3, although partial homology was observed to a number of sequences from EST databases. We designated this new clone NCU-G1. Northern hybridization experiments revealed that NCU-G1 is expressed constitutively not only in the mouse fetus but also in various mouse tissues, and is most abundant in the kidney cortex.

特発性縦隔気腫は比較的稀な疾患である.外傷等による続発性縦隔気腫を鑑別除外できれば, 通常治療は安静のみで良く, 数日中に軽快する.われわれの経験した症例は12歳男性で, サッカーの練習にてランニング中に突然の胸痛と呼吸苦感を訴え, 当科を受診した.頸部の皮下気腫と, 胸部X線及びCTにて縦隔内の気腫像を認め, 縦隔気腫の診断で入院となった.外傷による続発性縦隔気腫が否定できなかったため, 上部消化管内視鏡検査及び気管支鏡検査を行ったが, 異常所見を認めず, 特発性縦隔気腫の確定診断を得て, 安静, 予防的抗生剤投与の保存的治療にて治療を開始した.第4病日には, 胸部CT上縦隔気腫は完全に消失し, 第5病日に退院.その後半年経過した現在運動などの行動制限は行っていないが, 再発は認めていない.

Inflammatory pseudotumors of the bladder are rare in children. We describe a 7-day-old male neonate with inflammatory pseudotumor of the bladder. He presented with a 3-day history of macroscopic hematuria. Ultrasonography, computed tomography and cystoscopy showed an intravesical mass arising from the right lateral wall. Pathologic findings obtained by open excision revealed that the tumor had spindle-shaped cells without significant atypia infiltrating into submucosal fibrous tissue. There has been no evidence of recurrent tumor 12 months post-operatively. To our knowledge our case is the first presentation in a neonate among the reported pediatric cases.

Congenital vesicovaginal fistula is a very rare entity, the etiology of which has not been clearly elucidated because pathologic features have not been mentioned in previous reports. The case of a 4-year-old girl having incontinence resulting from a congenital vesicovaginal fistula joining with the left ectopic ureter from the hypoplastic kidney is described. This is thought to be the first presentation of congenital vesicovaginal fistula joining with ectopic ureter. A microscopic examination revealed the fistula consisting of transitional cell epithelium, suggesting an abnormal fusion of the ureteral bud and caudal end of the müllerian duct with the urogenital sinus.

A 6-year-old boy received renal transplantation and was treated with methylprednisolone, cyclosporine A and mizoribine. He developed Epstein-Barr virus-associated malignant lymphoma at 10 years and thyroid papillary carcinoma at 20 years of age. Chemotherapy for the malignant lymphoma was done after withdrawal of cyclosporine A and mizoribine, and thyroidectomy was performed for thyroid carcinoma. He was well and his serum creatinine was 1.0 mg/dl at 22 years of age. To our knowledge, no pediatric renal transplant recipient who had thyroid carcinoma or two different types of tumor has been reported in Japan.

We analyzed the presentation, treatment and survival of 4 children with renal cell carcinoma.We retrospectively reviewed the pathological and hospital records of 4 Japanese children diagnosed with renal cell carcinoma at our hospital from 1970 to 1998.In the 1 boy and 3 girls with an average age of 8 years 7 months at diagnosis the most common presenting complaints were gross hematuria in 75% and a palpable abdominal mass in 50%. Computerized tomography revealed characteristic calcification within the tumor in 3 of the 4 patients (75%). In the remaining case the lesion had high density areas with microcalcification, as confirmed by histopathological study. In 2 patients with regional lymph node metastasis calcification was also observed in the metastatic lesions. Disease was stages I to III in 1, 1 and 2 patients, respectively. All patients underwent transabdominal nephrectomy with regional lymphadenectomy. One patient with stage I disease had multiple metastases 15 months later and died of disease 55 months postoperatively. However, the remaining 3 patients received adjuvant interferon therapy and they are without evidence of recurrence a mean of 51.3 months postoperatively.Calcification within the tumor and/or metastatic lesions or high density areas in the tumor on screening computerized tomography are characteristic findings suggestive of pediatric renal cell carcinoma. Adjuvant therapy with interferon may provide some benefit in select pediatric patients. Further studies of a larger number of pediatric renal cell carcinoma cases may be necessary to establish the optimal diagnostic and therapeutic regimen.

The spiroketals containing a benzyloxymethyl moiety at the C8 position showed the most potent apoptosis-inducing activity, whereas its analogous compounds lacking any substituent at C8 or possessing ones other than the benzyloxymethyl moiety at C8 were all much less active. These results strongly suggest an important role of the benzyloxymethyl moiety linked to the C8 oxygen atom.

BACKGROUND Recent advance of perinatal ultrasound screening and/or physician's awareness of renal damage from recurrent pyelonephritis has brought about the increasing number of infants with primary vesicoureteral reflux (VUR) including refluxing megaureter which should be conceptually differentiated from simple high grade VUR. We evaluated the clinical outcome of infants diagnosed with refluxing megaureter. PATIENTS AND METHODS We retrospectively reviewed the clinical records of 15 infants (17 ureters) diagnosed as refluxing megaureter (max caliber > or = 10 mm) at our institution from 1988 to 1997. We compared the clinical outcome of refluxing megaureter with that of high grade VUR. (Results) Patients were 13 boys and 2 girls. Megaureter was unilateral in 13 patients and bilateral in 2. Fourteen infants (93.3%) presented with febrile urinary tract infection (UTI). The diameter of megaureter was 10-21 mm (average: 13.6 +/- 4.0 mm) at excretory urogram. Nine of 15 infants (60.0%) had breakthrough urinary infection. Its incidence was significantly higher than that of high grade VUR (21.3%) (p = 0.02). In 13 cases surgical treatments were performed, however 2 cases (max caliber: 16 mm, 21 mm) by Politano-Leadbetter or Paquin procedure required re-ureteroneocystostomy by Psoas-hitch procedure because of persistent reflux and recurrent UTI. On the other hand no patient required re-ureteroneocystostomy in high grade VUR. CONCLUSION It is important to differentiate refluxing megaureter from high grade VUR due to high incidence of breakthrough UTI. Ureteral remodeling and/or Psoas-hitch procedure are strongly recommended for adequate length of submucosal tunnel in refluxing megaureter.