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Fabry disease is an X-linked disorder of α-galactosidase A (GLA) deficiency. Our previous interim analysis (1 July 2014 to 31 December 2015) revealed plasma globotriaosylsphingosine as a promising primary screening biomarker for Fabry disease probands. Herein, we report the final results, including patients enrolled from 1 January to 31 December 2016 for evaluating the potential of plasma globotriaosylsphingosine and GLA activity as a combined screening marker. We screened 5691 patients (3439 males) referred from 237 Japanese specialty clinics based on clinical findings suggestive of Fabry disease using plasma globotriaosylsphingosine and GLA activity as primary screening markers, and GLA variant status as a secondary screening marker. Of the 14 males who tested positive in the globotriaosylsphingosine screen (≥2.0 ng/mL), 11 with low GLA activity (&lt, 4.0 nmol/h/mL) displayed GLA variants (four classic, seven late-onset) and one with normal GLA activity and no pathogenic variant displayed lamellar bodies in affected organs, indicating late-onset biopsy-proven Fabry disease. Of the 19 females who tested positive in the globotriaosylsphingosine screen, eight with low GLA activity displayed GLA variants (six classic, two late-onset) and five with normal GLA activity displayed a GLA variant (one classic) and no pathogenic variant (four late-onset biopsy-proven). The combination of plasma globotriaosylsphingosine and GLA activity can be a primary screening biomarker for classic, late-onset, and late-onset biopsy-proven Fabry disease probands.

The opossum Monodelphis domestica presents movement of the temporomandibular joint (TMJ) reflecting adaptation to eating habits similar to movement in humans, but the structure of the TMJ is not yet known. Thus, nine young M. domestica, of both sexes were weighed, anesthetized with xylazine (10 mg kg(-1) ), and ketamine (70 mg kg(-1) ) and processed for: 1. The analyses of the macroscopic angioarchitecture after latex injection, as well as the topography of the TMJ; 2. The analysis of microvascularization after injection of Mercox resin and corrosion of soft tissue with NaOH using scanning electron microscopy and; 3. The histological evaluation of the TMJ with an optical microscope. Macroscopic analysis of the latex injected vessels revealed the distribution of the arteries from the common carotid artery, receiving branches of the superficial temporal and maxillary arteries. The mandibular condyle has the long axis in the lateral-lateral direction, and is convex in the anterior-posterior direction. Its topography was determined in relation to the eye and external acoustic meatus. With scanning electron microscopy, microvascularization consists of arterioles of varying diameter (85-15 µm) of the meandering capillary network in the retrodiscal region, and a network of straight capillaries in the TMJ anterior region. Via light microscopy the TMJ has similar histological features to those of humans. These macroscopic, microscopic and ultrastructural data from TMJ of the M. domestica could be a suitable model for TMJ physiology and pathophysiology studies for then speculate on possible human studies. Microsc. Res. Tech. 79:806-813, 2016. © 2016 Wiley Periodicals, Inc.

High glucose (HG) induces production of transforming growth factor-beta1 (TGF-β1), but the mechanism remains elusive. The aim of this study was to determine the gene(s) involved in HG-induced TGF-β1 production in human peritoneal mesothelial cells (HPMCs). Microarray analysis was performed following a 3-h preincubation of HPMCs in 4 or 0.1 % glucose medium. Transcriptional genes were selected using Gene Ontology analysis for biological processes, including regulation of transcription and DNA-dependent. The effects of small interfering RNA (siRNA) treatments on the up-regulation of TGF-β1 mRNA were assessed by quantitative real-time polymerase chain reaction (qPCR). Finally, enzyme-linked immunosorbent assay (ELISA) was performed to determine which gene(s) contribute to the production of TGF-β1 protein in the medium. Microarray analysis revealed that the expression of 51 genes increased by more than 3-fold. Gene ontology analysis identified 13 genes for further study. qPCR confirmed mRNA amplification for 9 of the 13 genes. Furthermore, HG-induced up-regulation of TGF-β1 mRNA was attenuated by the siRNA of 4 genes: MDS1 and EVI1 complex locus (MECOM), FBJ murine osteosarcoma viral oncogene homolog B (FOSB), FBJ murine osteosarcoma viral oncogene homolog (FOS) and activating transcription factor 3 (ATF3). ELISA showed that siRNA treatment of FOS, but not MECOM, FOSB or ATF3, suppressed the increase of TGF-β1 protein in the medium. FOS is a downstream effector of HG stimulation in HPMCs that contributes to TGF-β1 production, suggesting that blocking FOS expression may be a therapeutic target for peritoneal fibrosis.

Chronic kidney disease patients share clinical and pathological features with the general aging population. Increased oxidative DNA damage, accumulation of cell cycle-arrested cells and decreased Klotho expression are assumed to be age-related factors that are reportedly linked to kidney disease. This study sought to determine the association between these age-related factors and renal damage in patients with IgA nephropathy (IgAN). We performed a cross-sectional analysis of 71 patients who were diagnosed with IgAN by renal biopsy. Expression of 8-hydroxydeoxyguanosine (8-OHdG, a marker of oxidative DNA damage), p16 (a marker of cell cycle-arrest) and Klotho (an anti-aging protein) were evaluated by immunohistochemical staining of renal biopsy samples. We correlated the changes in expression of these markers with Lee’s pathologic grades and the Oxford classification. We also investigated the independent association between these markers and interstitial fibrosis using multiple linear regression analysis. 8-OHdG and p16 increased but Klotho decreased with progression of pathologic grade. Expression of 8-OHdG and p16 increased with the deterioration of mesangial hypercellularity and segmental glomerulosclerosis. In addition, p16 increased but Klotho decreased with progression of tubular atrophy/interstitial fibrosis. In univariate regression analysis, age, body mass index, systolic blood pressure, urinary protein excretion and expression of 8-OHdG, p16 and Klotho showed significant correlations with interstitial fibrosis. Multivariable regression analyses revealed that aging, increased renal expression of p16 and decreased expression of Klotho were independently correlated with interstitial fibrosis. The age-related factors might play important roles in the development of IgAN.

Combination therapy of aliskiren and an angiotensin II receptor blocker (ARB) has been reported to be effective for reducing the level of proteinuria. However, it remains unclear whether this combination therapy contributes to suppression of kidney disease progression. The aim of this study was to investigate the effect of aliskiren on hard renal endpoints, when added to an ARB, in patients with advanced chronic kidney disease (CKD). The study design was a prospective, randomized open-label design. 83 CKD patients (52 men and 31 women) were enrolled and assigned randomly to an aliskiren add-on group (n = 42) or control group (n = 41). Entry criteria included elevated serum creatinine ≥1.5 mg/dl, urine protein excretion (≥1+ on urine dipstick test), and hypertension. All participants were treated with an ARB. The follow-up period was 12 months. 12 participants were withdrawn during the study period and the study was terminated in January 2012 as a consequence of the results of the interim analysis of the ALTITUDE study. Nine patients in the aliskiren group and seven patients in the control group started dialysis. Doubling of the serum creatinine level occurred in one patient in the control group. A Cox proportional hazards test showed that dual blockade of the renin–angiotensin–aldosterone system with aliskiren and ARB was not associated with improvement in hard renal endpoints. We conclude that aliskiren add-on therapy to an ARB may not give any benefit and, therefore, should not be recommended in CKD patients.

This paper presents a low-capacity cation-exchange chromatography method for the analysis of UV-absorbing dipeptides and amino acids. A newly marketed low-capacity cation-exchange column packed with sulfo-functionalized highly cross-linked macroreticular poly(ethylstyrene-divinylbenzene) copolymer was used for the simultaneous determination of imidazole amino acids, aromatic amino acids, and creatinine in urine samples. A dual-mode binary gradient chromatography method was established using two solvents, A: 15 mM H3PO4/5 mM ethylenediamine and B: 15 mM H3PO4/5 mM ethylenediamine/40 (v/v) % CH3CN at 40 °C, with an optimized time program for changing the delivery ratio of A/B and the flow rate. Good chromatograms were obtained within an acceptable cycle time of 25 min. The quantification data were satisfactory for all analytes, showing the relative standard deviations (RSD) of retention times between 0.08 and 1.68 %; RSDs of area intensities between 0.23 and 2.60 %; and linear regression lines with r 2 more than 0.9994. The method could determine the creatinine ratios of the diagnostic markers on the single chromatographic run, which enabled to discriminate disease from health. For example, the creatinine ratios for phenylketonuria were significantly higher than those for controls. The method can provide highly cost-efficient information or useful knowledge for clinical and pharmaceutical studies.

Mesenchymal stem cells (MSCs) are multipotent adult stem cells that have regenerative capability and exert paracrine actions on damaged tissues. Since peritoneal fibrosis is a serious complication of peritoneal dialysis, we tested whether MSCs suppress this using a chlorhexidine gluconate model in rats. Although MSCs isolated from green fluorescent protein–positive rats were detected for only 3 days following their injection, immunohistochemical staining showed that MSCs suppressed the expression of mesenchymal cells, their effects on the deposition of extracellular matrix proteins, and the infiltration of macrophages for 14 days. Moreover, MSCs reduced the functional impairment of the peritoneal membrane. Cocultures of MSCs and human peritoneal mesothelial cells using a Transwell system indicated that the beneficial effects of MSCs on the glucose-induced upregulation of transforming growth factor-β1(TGF-β1) and fibronectin mRNA expression in the human cells were likely due to paracrine actions. Preincubation in MSC-conditioned medium suppressed TGF-β1-induced epithelial-to-mesenchymal transition, α-smooth muscle actin, and the decrease in zonula occludens-1 in cultured human peritoneal mesothelial cells. Although bone morphogenic protein 7 was not detected, MSCs secreted hepatocyte growth factor and a neutralizing antibody to this inhibited TGF-β1 signaling. Thus, our findings imply that MSCs ameliorate experimental peritoneal fibrosis by suppressing inflammation and TGF-β1 signaling in a paracrine manner.

The opossum Monodelphis domestica presents movement of the temporomandibular joint (TMJ) reflecting adaptation to eating habits similar to movement in humans, but the structure of the TMJ is not yet known. Thus, nine young M. domestica, of both sexes were weighed, anesthetized with xylazine (10 mg kg(-1) ), and ketamine (70 mg kg(-1) ) and processed for: 1. The analyses of the macroscopic angioarchitecture after latex injection, as well as the topography of the TMJ; 2. The analysis of microvascularization after injection of Mercox resin and corrosion of soft tissue with NaOH using scanning electron microscopy and; 3. The histological evaluation of the TMJ with an optical microscope. Macroscopic analysis of the latex injected vessels revealed the distribution of the arteries from the common carotid artery, receiving branches of the superficial temporal and maxillary arteries. The mandibular condyle has the long axis in the lateral-lateral direction, and is convex in the anterior-posterior direction. Its topography was determined in relation to the eye and external acoustic meatus. With scanning electron microscopy, microvascularization consists of arterioles of varying diameter (85-15 µm) of the meandering capillary network in the retrodiscal region, and a network of straight capillaries in the TMJ anterior region. Via light microscopy the TMJ has similar histological features to those of humans. These macroscopic, microscopic and ultrastructural data from TMJ of the M. domestica could be a suitable model for TMJ physiology and pathophysiology studies for then speculate on possible human studies. Microsc. Res. Tech. 79:806-813, 2016. © 2016 Wiley Periodicals, Inc.

A research project was conducted in which proper JAPAN/MARC bibliographic records for 158 major Japanese classical works were identified manually, since existing records contain little information about works included in the resources. This article reports on the detailed method used for work identification, including selecting works, obtaining the bibliographic records to be judged, and building the judgment criteria. The results of the work identification process are reported along with average numbers that indicate the characteristics of certain classics. The necessity of manual identification was justified through an evaluation of searches by author and/or title information in a conventional retrieval system.

When diagnosing hypertension (HT) it is essential to determine not only the level of raised blood pressure (BP), but also how the condition relates to organ damage. The best time to measure BP for diagnosing HT in patients on hemodialysis (HD) remains unclear. A total of 100 HD patients (mean age 63.8 years, 60 males) were studied. Left ventricular hypertrophy (LVH) was detected by echocardiography and BP monitored for 1 week at 20 different times in the morning and night, before and after dialysis. We also checked for masked HT, i.e., patients with weekly morning HT, but not pre-dialysis HT. Average BP for the week was 141.9 ±19.0/79.6 ± 10.6 mmHg, with 68 patients classified as hypertensive. Average morning BP was 144.6 ± 19.8/81.7 ± 11.3 mmHg, and 71 patients had weekly morning HT. In addition, 62 patients had LVH and 51 patients had relative morning HT. Multiple logistic analyses showed that LVH was associated with weekly morning HT, morning HT on HD and non-HD days, average HT, and relative morning HT. However, evening, pre-dialysis, and post-dialysis HT showed no association with LVH. Masked HT was found in 20 % of patients. If HT had been diagnosed using only pre-dialysis BP, 20 of the 71 patients with weekly morning HT would not have been detected. Morning BP is useful for detecting LVH in HD patients. Monitoring of morning BP may be superior to measurements taken at other times for diagnosing HT.

A possible formation mechanism of the ionic aggregate of poly(oxyethylene) dodecyl ether nonionic surfactants with a ferric-thiocyanate chromophore is presented, and the dependence of their colorimetric sensitivity on the degree of ethoxylation can be clearly explained through both experimental and theoretical approaches. The stoichiometry of ferric thiocyanate was confirmed to be a monovalent anion comprised of the 1:4 complex, [Fe(SCN)4]−, both by the extractive mole-ratio method and the continuous variation method. The poly(oxyethylene) moiety can form a stable helical conformation by including an alkali cation inside the helix, leading such nonionic surfactants into cationic complexes that can produce ionic aggregates with a bulky anionic complex, such as ferric thiocyanate. The colorimetric sensitivity of the ionic aggregates can depend on the inter-charge distance between the cation and the counter anion. The newly introduced conformational ideas based on experimental results are theoretically supported by CHARMM29 molecular mechanics/dynamics simulations. Molecular dynamics simulations successfully provided the most stable conformation for the K+-included heptaoxyethylene dodecyl ether aggregated with the bulky counter anion.

The analysis or preparative isolation of a specimen by a packed column gas chromatograph is affected by the kind and amount (concentration) of the liquid phase coated on the stationary phase. In particular, compounds having the same or similar molecular weight and similar functional groups are largely affected. Methyl oleate, which is a typical component of fats and oils, produces hydroperoxide by air oxidation as a first step. After that, various oxygenated compounds with a carbon number of 19 are produced. In this study, the effect of the concentration of ethylene glycol succinate polyester (EGS) as the liquid phase was examined for these compounds. The retention time of the hydroxides increased with an increase in the EGS concentration. The effect of the EGS concentration on the retention time decreased when these compounds were trimethyl silylated (TMS). The retention time of epoxides and oxo compounds increased with an increase in the EGS concentration independently of trimethyl silylation. Correction values were required for all quantitation.

A highly sensitive spectrophotometric determination of cationic surfactants in ground waters was established by forming their Cu(II)–TPPS aggregates, preceded by solid-phase extraction with an SCX cartridge. Cationic surfactants (CSs) were quantitatively trapped and isolated by the SCX solid phase. The use of Cu(II)–TPPS anionic chromophore could reduce the interference by unintentional metal ions coexisting in surrounding waters. The method was very sensitive in the determination of CSs less than 10 −5 M levels with acceptable recovery and calibration data. The colorimetric sensitivity was very dependent on the alkyl-chain length of the surfactants, and a CS having 23 carbon atoms gave the highest sensitivity. Overall recoveries were 95–97% with R.S.Ds. less than 3% in the cases over 10 −6 M levels. In the cases in 10 −7 M levels, however, a portion of the analyte would be adsorbed by reservoir walls, which could seriously affect the trace determination. The preliminary addition of 4,4′-bipyridyl into the sample solution was effective in decreasing such unintentional analyte losses, leading up to 73% recovery. The developed method was applied to the analysis of river water at ppb levels of CSs with a fractional concentration through a SCX solid phase subjected by 500-mL aliquots of sample.

SUMMARY: Background: In vitro studies suggest that the signal transducer and activator of transcription (STAT) plays a critical role in renal fibrosis. However, the process of STAT activation in vivo remains unclear. This study in rats aimed to localize STAT3 activation within the kidney and examine the in vivo relationship between STAT3 activation and renal fibrosis. Methods: Unilateral ureteral obstruction (UUO) was induced in the rats and the kidneys examined 3 or 7 days after obstruction. Activation of STAT3 in western blot and immunohistochemical analyses was identified by the phosphorylated form of STAT3 (pSTAT3). Results: Myofibroblasts were identified by α-smooth muscle actin expression and were upregulated in obstructed kidneys. pSTAT3 was localized mainly in tubular epithelial cells of collecting ducts in normal and obstructed kidneys and interstitial cells in obstructed kidneys. After UUO, western blotting showed a fourfold increase in pSTAT3, with a peak at day 7. Immunostaining showed a sixfold increase in pSTAT3 at day 7 in tubular epithelial cells and a 2500-fold increase at day 7 in interstitial cells. Conclusion: STAT3 was activated in rat tubular epithelial cells and myofibroblasts after UUO, suggesting that STAT3 may contribute to the progression of interstitial fibrosis.

Relative alkali-cation affinity of polyoxyethylene (POE) dodecylethers in gas phase was studied by electrospray ionization (ESI) mass spectrometry using dodecylether-poly-ethoxylate (C(12)EO:n, "n" denotes ethyleneoxide unit number) nonionic surfactants, and possible helical conformations of the cationized molecules were demonstrated. The alkali-cation affinity highly depended on the cation diameters. The mass spectra of C(12)EO:8 cationized by alkali-metal ions were dominated by potassiated molecules. The results indicated that the POE moiety could have specific affinity to K(+) ions based on a host-guest interaction between POE helix and potassium ions. This is very similar to the relationships between 18-crown-6 and K(+). The ESI mass spectra exhibited the multiply cationized C(12)EO:n in addition to the singly cationized molecules. The critical EO unit numbers necessary for producing the multiply-charged cationized molecules also depended on the cation diameters. In addition, the POE surfactants highly preferred alkali cations to proton. The results were strongly supported by molecular mechanics/dynamics calculations. A helical conformation of the POE moiety of C(12)EO:15 including two K(+) ions gave a potential minimum, while a lowest energy structure of the protonated molecule took irregular conformations due to the formation of local hydrogen bonds.

【目的】がんの在宅緩和ケアを支える手段として既存の日本のデイケア施設がいわゆる英国的デイホスピスとして機能しうるかどうかを知るために, デイケア施設におけるがん患者に対する医療サービスの実態について調査したので報告する. 【対象と方法】2004年11月岡山県のデイケア108事業所に対してアンケートを行った. 【結果】現在がん患者の利用がある事業所は56%, がん患者のための特別なプログラムを有している事業所は8%あった. デイケア施設が在宅緩和ケアを支える場合に備えるべき機能として, 1)がんの緩和医療に必要な医療サービス2)医療監視下における入浴等のデイケアサービス3)がん患者のニーズにあわせた休息を取り入れたサービス4)ストマケア, IVH管理, 輸液等のがん支持療法としての医療機能5)患者・家族に対する精神的ケア等が考えられた. 【結論】日本のデイケア施設において英国的緩和デイケアサービスを提供することは可能と思われた.

This paper presents a new HPLC technique for the determination of biogenic cations such as amino acids and nucleobases, using a weak-acid cation-exchange column. Fourteen analytes, five amino acids and seven bases in addition to creatinine and creatine, were separated in 12 min by means of a two-liquid gradient elution with UV detection. The newly released column packed with a carboxy-functionalized polymethacrylate resin could give excellent selectivity to the organic cations of interest, although such a column is in general suitable for the separation of inorganic common cations. The chromatographic intra-day repeatability was very good with RSDs less than 0.4%, and the quantitation precision based on peak area intensities was also good with RSDs less than 5% for all analytes. The linear calibration lines for quantitation ranged between 5 and 500 μM on 20-μL injections with R(2) more than 0.9990. Since the method could provide concentration data of urinary creatinine and some metabolites simultaneously, for example, the urinary phenylalanine/creatinine ratios for phenylketonuria of inborn errors of metabolism were simply determined through one chromatographic run. The ratios for patients were significantly higher than those for controls. We found that the new weak-acid cation-exchange column was suitable for the separation of organic cations as well as inorganic cations.

Objective Although lipid disorders are a well-known risk factor for cardiovascular disease (CVD) in the general population, the optimal management with lipid-lowering therapy to reduce CVD risks and mortality in hemodialysis (HD) patients remains controversial. In the clinical setting, dyslipidemia can be diagnosed based on the detection of elevated lipid concentrations at the beginning of HD. This study investigated changes in the levels of serum lipids during a single HD session. Methods The serum total cholesterol, triglyceride and high-density lipoprotein (HDL) cholesterol levels were measured in 31 HD patients at zero, two and four hours after the beginning of a single HD session. The data were analyzed using the Wilcoxon signed-rank test, a linear mixed model and Spearman's rank correlation analysis. Results The serum total cholesterol, HDL cholesterol and non-HDL cholesterol levels increased significantly during the HD session. Even after the lipid parameters were corrected for changes in the total protein level, the total cholesterol and HDL cholesterol levels increased, whereas the non-HDL cholesterol levels did not change significantly. The percentage change in the serum levels of these lipid fractions correlated strongly with the percentage change in the ultrafiltration volume per body weight. In contrast, the serum triglyceride levels were decreased significantly at two hours compared with the levels noted at the beginning of HD and gradually increased at four hours. Conclusion The serum lipid levels are influenced significantly by HD treatment and ultrafiltration. Evaluating the degree of dyslipidemia at the beginning of a HD session may therefore underestimate the levels of serum lipids in HD patients with a large amount of weight gain, thus resulting in the use of insufficient lipid-lowering therapy.

サプレッサー付電気伝導率検出方式のイオンクロマトグラフで陰イオンを測定する場合の検量線の直線性について検討した．解離平衡に基づく計算シミュレーションで理論的な検量線を導出するとともに，弱塩基性及び強塩基性溶離液を用いてF－，Cl－，NO3－及びSO42－の検量線データを求めた．計算シミュレーションと実験結果を比較したところよい一致を示し，弱塩基性溶離液を使う場合，また，試料成分が弱酸の共役塩基である場合には，濃度範囲によって，また望む精度によって直線では十分近似できないことを明らかにした．また，理論上は検量線について高次の検量線を想定できても，広範囲の濃度では測定精度が異なるため，範囲を限定して1次で近似したほうがよいことを明らかにした．高精度測定のためには同一精度と見なすことのできるデータ群に限定して検量線の次数を検討する必要がある．

Commercially available but completely unknown surfactants used in the tin–lead plating industry were successfully identified by using electrospray mass spectrometry and NMR spectroscopy techniques, preceded by liquid ionization mass spectrometry used to obtain the preliminary information. The mass spectral data suggested that ethoxylated nonionic surfactants having a homologous distribution of molecular weights like 520, 564, 608, 652, 696, etc. were present. The NMR data suggested the presence of two aromatic rings and a quaternary carbon for the hydrophobe moiety instead of the well-known alkyl chains or alkylphenols. The unknown surfactants were finally concluded to be novel nonionic 4-(α,α-dimethylbenzyl)-phenol ethoxylates.

and ESI, in which [M + H]+ were not observed or the spectra were complicated. The liquid ionization mass spectra of triorganotin carboxylates varied with solvents and sample concentrations. For instance, the fragment ions [M + (C4H9)3Sn]+ of dimeric ions were observed with chloroform used as a solvent, while the [M + H]+ were observed as the base peak using ethylene dichloride. Spectra useful for the differentiation of isomers [CgH7O3Sn(C4Hg)3] were obtained by the formation of characteristic adduct ions, such as [M + EA + H]+ and [M + 2EA + H]+, with a reagent like 2-aminoethanol. Collision-induced dissociation (CID) spectra observed by ESI and LPI mass spectrometry were similar and provided less information than adduct ions did.

ポリオキシエチレン系非イオン界面活性剤(NS)のチオシアン酸鉄(III)錯体形成を用いる抽出吸光光度定量法において,操作に使用する試薬量の低減化を図った。その結果,従来法に比べて約1/5のスケールダウンを実現でき,従来法とほとんど同じ定量結果が得られることが分かった。NS標準物質としてヘプタオキシエチレンドデシルエーテルを用いて検量線を作成した結果,濃度0.1～2.0mg/lの範囲で吸光度との間に良好な直線関係が得られた。JIS法に比べ約24倍の感度が得られ,使用する試薬の量もJIS法と比較して大幅に減らすことができた。但し,本法も,NSの抽出吸光光度定量において従来から一般的に指摘されているように,イオン性界面活性剤,特に陽イオン界面活性剤が存在すると正誤差を与え,また検出感度はNS分子中のオキシエチレン鎖長に依存することも確認された。

症例は68歳, 男性.右肺尖部の腫瘤影を指摘されたが放置.約半年後に腫瘤影の増大を認めたため当院内科に入院となった.右肺S2原発のPancoast型肺癌 (cT3N2MO) の診断にて, ドセタキセル・シスプラチン併用療法を2クール施行したが治療効果は「不変」であった.外科転科後, 右肺上中葉切除および胸壁合併切除術を施行した.組織型は低分化型腺癌であった.術前N2と判定された縦隔・肺門リンパ節の腫大は珪肺によるものと判明し, 術後病期はpT3NOMO (旧分類IIIA期, 新分類IIB期) であった.本患者の末梢血中CEA mRNA検出をRT-PCR法にて行ったところ, 術前および術後15日目の末梢血ともにCEA mRNAは陽性であった.

The effects of amines on the formation of molecular adduct ions of methyl hydroperoxyoleates (OMH) in atmospheric pressure chemical ionization mass spectrometry were investigated. The short-chain primary amines such as methylamine and ethylamine were more effective than ammonia on suppressing the thermal decomposition of OMH, and provided abundant [M+H+L]+ type ions (L: each amine molecule). The secondary and tertiary amines were unsuitable for the purpose in spite of their high basicity, probably due to the steric interference. Aromatic and heterocyclic amines were also ineffective. The formation of amine adduct to OMH in liquid and/or condensed phases depends on both basicity and stereochemistry of each amine.

The method for observing molecular related ions of methyl hydroperoxyoleates (OMH) by atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) was investigated. Since thermally labile OMH is decomposed in the APCI nebulizer and/or desolvation chamber heated to a high temperature, it is difficult to observe significant peaks of their protonated molecules. The addition of ammonia water to the mobile phase (ethanol) was quite effective to suppress the thermal decomposition of OMH. As the result the mass spectra exhibited the intense peaks of ammonium adduct ions of OMH, [M+NH4]+, at m/z 346. The nebulizer temperature at 170°C and the drift voltage at 170 V were adequate for observing the molecular ion species of OMH.

An optimum combination of reversed-phase column type and eluent composition was selected for the practical separation of amino acids by gradient elution ion-pair chromatography. A butyl-silica (C4) column with a concentration gradient elution between 5mM SDS-10mM HClO4 solution and acetonitrile provided the best resolution for amino acids. The C4 column was most adequate for reducing the chromatographic cycle time including the column regeneration. The use of 5mM SDS as ion-pairing agent in relatively low concentration is economical and practical for replicate and routine analyses. The developed HPLC system is applicable to the analysis of organic cations at low cost.

Aldosterone-salt treatment induces not only hypertension but also extensive inflammation that contributes to fibrosis in the rat kidney. However, the mechanism underlying aldosterone-salt-induced renal inflammation remains unclear. Pyroptosis has recently been identified as a new type of cell death that is accompanied by the activation of inflammatory cytokines. We hypothesized that aldosterone-salt treatment could induce inflammation through pyroptosis and that mizoribine, an effective immunosuppressant, would ameliorate the renal inflammation that would otherwise cause renal fibrosis. Ten days after recovery from left uninephrectomy, rats were given drinking water with 1% sodium chloride. The animals were divided into three groups (n = 7 per group): (1) vehicle infusion group, (2) aldosterone infusion group, or (3) aldosterone infusion plus oral mizoribine group. Aldosterone-salt treatment increased the expression of the nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing 3 and caspase-1, and also increased the number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells. However, the oral administration of mizoribine attenuated these alterations. Furthermore, mizoribine inhibited hypertension and renal fibrosis, and also attenuated the aldosterone-induced expression of serum/glucocorticoid-regulated kinase and α epithelial sodium channel. These results suggest that caspase-1 activation plays an important role in the development of inflammation induced by aldosterone-salt treatment and that it functions as an anti-inflammatory strategy that protects against renal injury and hypertension.

A general, functional relationship between the retention volume of the 2nd system peak and the chromatographic parameters (the acid dissociation constant of the eluent component, the eluent concentration, the acid/salt ratio in the eluent, the acid-partition coefficient) in ion chromatography with monobasic acid eluent systems (acids, salts, and their mixtures) have been derived. The relations precisely reproduced the results by computer simulation based on the plate theory, and were confirmed with independent experiments. By using these relations, the partition isotherm of eluent acids to the stationary phase can be obtained easily.

A non-suppressed ion chromatographic (IC) system for the high-sensitivity detection of common anions was developed using sulfoisophthalic acid as the eluent. The detection sensitivity was ten times higher than that using conventional non-suppressed IC with sodium phthalate as eluent, and was almost the same as that using conventional suppressed IC with a carbonate-hydrogencarbonate eluent under the same electrical conditions with a conductivity detector. Temperature regulation was very important in minimizing the baseline drift. A commercial incubator, in which a separation column and a sample injector were placed, was useful. The developed non-suppressed system facilitated the determination of low concentrations of phosphate, chloride, bromide, nitrate and sulfate at micromolar levels.

Effects of additives, introduced to a sample through vapor phase, on the formation of protonated molecules and/or molecular adduct ions were investigated in liquid ionization mass spectrometry for the analysis of methyl hydroperoxyoleates. Water and formic acid having small proton affinities were useful for increasing ion abundance of protonated molecules [M+H]+. 2-Aminoethanol and pyridine having slightly greater proton affinities were useful for forming molecular adduct ions [M+H+L]+ (L: additive molecule). The use of additives, acting as matrix for producing [M+H]+ and/or as reagent for forming [M+H+L]+ in liquid ionization mass spectrometry, provided useful mass spectral characteristics of the molecular weight and structure of unknown compounds.

High rate of growth are expected in the Pulp and Paper Industry of Indonesia, Thailand, China and Vietnam. These countries have great potentiality of becoming the most important area for the further development of Japanese Pulp and Paper Industry in the next century.The growth mainly result from the high rate of economic growth in these countries and various factors peculiar to each country.

A polyfunctional low-capacity cation-exchange column-packing material was developed for the simultaneous separation and determination of underivatized 20 amino acids including 16 proteinogenic ones using a binary gradient HPLC. The new packing material was prepared by sulfoacylating a highly crosslinked macroreticular poly(styrene-divinylbenzene) copolymer of 3 μm in diameter commercially available for size-exclusion chromatography. The polyfunctionality could derive from unintentional carboxy groups innately present in the base polymers probably came from polymerization initiators in addition to the intentionally introduced sulfopropionyl groups, which was certified and evaluated by measuring dynamic capacity curves of the cation exchanger. Sixteen underivatized proteinogenic amino acids were separated in 23 min using a binary high-pressure gradient elution system with two liquids of 1 mmol L(-1) H(3)PO(4) and 20 mmol L(-1) NaH(2)PO(4)/30% (v/v) CH(3)CN, which led to the cycle time of 25 min. In the case of separation of 20 amino acids, the cycle time was 27 min. The developed low-capacity cation-exchange chromatography with post-column fluorescence detection was sufficiently quantitative, providing linear calibration lines ranged through almost three digit for all analytes. The detection limits were calculated as nmol L(-1) order of magnitude with 20-μL injection. The method was applicable to the direct analysis of urinary amino acids of diagnostic markers for inborn errors of metabolism.

A mixture of isomeric methyl 9-, 10-, 11-, and 12-hydroperoxyoleates was analyzed by liquid ionization (LI) mass spectrometry. LI mass spectra of the hydroperoxides exhibited [M+H]+ peaks clearly at m/z 329, while both chemical ionization and fast atom bombardment mass spectra did not show ions directly related to the molecular weight. The fragment peaks of the hydroperoxides observed at m/z 295, corresponding to the loss of H2O2 from [M+H]+, were useful to discriminate the hydroperoxides from structurally isomeric hydroxyepoxides. In addition, several characteristic fragments were observed at m/z 173, 187, 199, 201, and 213, which were also indicative of the position of the hydroperoxy group in the hydroperoxides.

A spectrometric method for the determination of anionic surfactants in environmental water samples using the [Co(III)-(5-Cl-PADAP)(2)](+) chromophore was technically improved for practical use. An anisole-extraction procedure was alternatively introduced and optimized in place of the earlier benzene-extraction procedure. The molar absorptivity for SDS found by the Co(III)-5-Cl-PADAP/anisole method was 72000 L mol(-1) cm(-1) at 560 nm, which was ca. 1.3-times higher than those by the Co(III)-5-Cl-PADAP/benzene method. The analytical sensitivity was almost independent of the kind of anionic surfactants. The calibration lines for SDS, LAS, and SSS were all linear between 1.0 × 10(-7) and 1.0 × 10(-5) mol L(-1) with acceptable r(2) values of 0.9998, 0.9995, and 0.9999, respectively. The method was applied to the analyses of river, well, and seawater samples, providing the compatible results by means of the standard-addition method. In the case of seawater, a phase-washing treatment of the extracted species using dilute HCl was effective to decrease the high blank absorbance. The Co(III)-5-Cl-PADAP/anisole method, showing no glass-adsorption problem, is more advantageous than the methylene-blue and ethyl-violet methods.

We investigated whether or not N-terminal pro brain natriuretic peptide (NT-proBNP) could predict hospitalization for cardiovascular disease (CVD) among Japanese hemodialysis patients. A total of 104 patients on maintenance dialysis 3 times per week were enrolled. We followed the patients for 23.9 +/- 4.2 months and 19 hospitalizations for CVD occurring during this period. The area under the curve (AUC) for the risk of CVD hospitalization was calculated after drawing a receiver operating characteristic curve. Predialysis NT-proBNP showed a larger AUC value than both postdialysis NT-proBNP and brain natriuretic peptide. The optimal cut-off value of predialysis NT-proBNP for predicting CVD hospitalization was 5,894 pg/mL, (sensitivity of 60 % and specificity of 76 %). Diabetes mellitus, a history of CVD, and the predialysis NT-proBNP level were significant determinants of CVD hospitalization according to Cox proportional hazards analysis. In conclusion, predialysis NT-proBNP is useful for predicting CVD hospitalization in hemodialysis patients.

Since activated factor X (FXa) is a coagulant enzyme that generates thrombin and participates in both intrinsic and extrinsic coagulation pathways, inhibition of FXa may be more effective than inactivation of thrombin for interrupting blood coagulation. To assess the possible effectiveness of FXa inhibition as an anticoagulant, we designed and synthesized 3-(amidinoaryl)-2-[4-[(3S)-3-pyrrolidinyloxy]phenyl]propanoi c acid derivatives as low molecular weight, nonpeptidic, orally active FXa inhibitors. These derivatives exhibited potent and highly selective anti-FXa activity in vitro and anticoagulant activity on oral administration. The most promising compound, (2S)-2-[4-[[(3S)-1-acetimidoyl-3-pyrrolidinyl]oxy]phenyl]- 3-(7-amidino-2-naphthyl)propanoic acid hydrochloride pentahydrate (4,DX-9065a), inhibited 50% of FXa activity (IC50) at 0.07 microM, doubled plasma recalcification time (PRCT) at 0.5 microM, and significantly prolonged activated partial thromboplastin time (APTT) at a dose of 100 mg/kg on oral administration. In contrast with FXa inhibition, 4 showed no activity against thrombin (IC50 > 2000 microM).

15-Deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) is an endogenous peroxisome proliferator-activated receptor gamma (PPARgamma) agonist that suppresses progressive matrix deposition; however, little is known about the effects of 15d-PGJ(2) on human peritoneal mesothelial cells (HPMCs). We investigated the following: (i) the expression of PPARgamma; (ii) the effect of 15d-PGJ(2) on angiotensin II (Ang II)-induced fibronectin (FN) expression and secretion; (iii) the effect of 15d-PGJ(2) (with or without Ang II and with or without the specific PPARgamma antagonist GW9662) and pioglitazone, a synthetic PPARgamma agonist, on hepatocyte growth factor (HGF) expression and secretion; (iv) the effect of HGF on Ang II-induced FN expression and secretion; (v) the expression of c-Met (a specific HGF receptor) and its phospho-signal; and (vi) the involvement of HGF in the effect produced by 15d-PGJ(2) using selective c-Met inhibitor PHA-665752. The presence of PPARgamma was detected by western blot analysis. 15d-PGJ(2) inhibited Ang II-induced FN expression and increased HGF expression, even in the presence of Ang II. This effect of HGF expression was completely prevented by co-treatment with GW9662. Additionally, upregulation of HGF secretion induced by 15d-PGJ(2) and HGF production induced by pioglitazone was revealed. We demonstrated the presence of c-Met, and presented evidence that HGF inhibits Ang II-induced FN expression and activates phosphorylation of c-Met, which is blocked by PHA-665752; 15d-PGJ(2) also activated c-Met phosphorylation. Furthermore, PHA-665752 attenuates the inhibitory effects of 15d-PGJ(2) on FN secretion. These findings suggest that 15d-PGJ(2) has a novel and potent antifibrotic effect in HPMC and this action is likely mediated by HGF.

The effective ion-exchange capacities of ion-exchange materials were determined by measuring the change in the equilibrium conductivity of a column packed with analyte. The developed instrumental method can provide effective ion-exchange capacities for both cation and anion exchangers with simple operations. The cation-exchange capacity of a weak-acid cation-exchange resin (TSKgel SuperIC-Cation column) depended on the conditioning pH and the molar concentration of the conditioning agent. Plots of effective cation-exchange capacities over the conditioning pH exhibited three inflection points, suggesting the presence of two carboxy groups and one phenolic OH group in the resin, probably due to the inherent base polymer. This method was applied to several commercial analytical columns for ion chromatography, and could provide scientifically useful results for characterizing the resin properties.

Peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand-dependent transcription factor that has a central role in the regulation of insulin sensitivity and adipocyte differentiation. Expression of PPARgamma has been reported in the kidney, including medullary collecting ducts, glomeruli and tubular cells. Thiazolidinediones (TZDs) are synthetic PPARgamma agonists and are used widely in patients with type 2 diabetes. It has been gradually discovered that TZDs have various other actions, such as vascular protective, anti-inflammatory, anti-fibrotic and anti-proliferative actions, over and above their effects on glucose and lipid metabolism. In this review, we will focus on current knowledge and insights on the role of PPARgamma agonists in kidney diseases, especially in diabetic nephropathy, non-diabetic kidney diseases and dialysis therapy.

A highly sensitive HPLC method for the simultaneous determination of soluble silicate and phosphate in environmental waters was developed, using ion-pair liquid chromatography preceded by the formation of their yellow alpha-heteropolymolybdates. The moderate-pH mobile phase enabled to use a highly efficient reversed-phase silica column. The pre-column coloring reactions at moderate-pH were reproducible for both silicate and phosphate in all quantification ranges with R.S.D.s less than 2% and 5%, respectively. The linear calibration lines between concentrations (mg-SiO(2)/L and mg-PO(4)/L) and peak area intensities were obtained for silicate and phosphate both with acceptable determination coefficients (r(2)) of 0.9999. The limits of determination for both analytes were 0.007 mg-SiO(2)/L and 0.003 mg-PO(4)/L, which were calculated theoretically using 10sigma/slope. The four-digit dynamic ranges were obtained for 0.007-10mg-SiO(2)/L and 0.003-20mg-PO(4)/L. The developed method was applied for the analysis of tap water, river water, coastal seawater, well water, hot-spring water, commercial mineral water, and laboratory water. The results were very reasonable and acceptable from the environmental viewpoints, which were well correlated with those confirmed by the molybdenum-blue spectrophotometry.

Phospholipids in middle ear effusion (MEE) and serum were analysed by liquid ionization (LI) mass spectrometry. The LI mass spectra of MEE and serum measured directly without preliminary separation commonly exhibited several unknown peaks such as those at m/z 520 and 577. These peaks were also observed in 90% ethanol fractions of MEE. The mass spectrum of the 90% ethanol fraction of serum exhibited dominant peaks at m/z 551, 577 and 603 (with mass differences of 26) and also at m/z 520. Comparing this spectrum with the mass spectra of standard compounds, the ions of mass 551, 577 and 603 were identified to be [M — phosphocholine] + of dipalmitoyl, palmitoyl-oleoyl and dioleoyl-glycero-3- phosphocholine (PC), respectively. The relative abundances of these ions observed in serum were qualitatively consistent with the composition of the fatty acid residue of PCs in serum determined by gas chromatography. The ions of mass 520, observed to be more abundant in MEE than in serum, were also identified to be [ceramide + — O], which is equal in mass to [M — phosphocholine] + , of N -palmitoyl-sphingo-phosphocholine (Sph). The results strongly suggest that a part of blood permeates into the middle ear cavity, and especially Sph does this easily. LI mass spectrometry is useful for analysis of phospholipids in biological samples.

A high-performance liquid chromatography method for the determination of linear alkylbenzenesulphonates (LASs) in river waters has been developed. The ppb levels of LASs can be determined by reversed-phase high-performance liquid chromatography with ultraviolet detection after on-line anion-exchange concentration and successive injection. LASs were quantitatively concentrated on the anion-exchange precolumn and easily cleaned up from river water matrix, because of its specific affinity, for the anion-exchange resin. A weak non-polar reversed-phase column was useful for the determination of LASs. The relationships between concentration andsummation of peak areas were linear from 10 to 200 ppb for total LAS concentrated from 5 ml of standard solutions. Overall recovery for total LAS was found to be 99%. Total LAS in the Tama River waters was determined to be around 100 ppb by the proposed method.