We propose here two new quantitative analytical methods for the determination of total nitrogen in environmental water samples. Firstly, a downsized alkaline peroxodisulfate digestion procedure for total nitrogen was proposed. An aliquot of the digested sample (the supernatant liquid) was introduced into a flow injection analysis (FIA) system in which the chemistry relied on a diazotization/coupling reaction with a copperized cadmium reduction column. The amounts of sample and digestion solutions were reduced to one-tenth and one fifth, respectively, compared with the present Japanese Industrial Standard (JIS) method. Furthermore the batchwise digestion procedure was able to treat not only clear (transparent) samples, but also samples including suspended solids so that various kinds of samples can be introduced into the subsequent FIA system. Secondly, we developed an on-line digestion FIA method for total nitrogen based on both of ultraviolet spectrophotometry of nitrate and the above mentioned diazotization spectrophotometry of nitrite. In the proposed FIA system equipped with both an ultraviolet and visible detectors, an injected sample solution was subject to on-line alkaline peroxodisulfate digestion, the absorbance of nitrate, itself, at 220 nm was measured, and then the absorbance of the azo dye at 540 nm was measured. In conclusion, the these two quantitative analytical methods were both applied to natural water analysis, and the analytical results were in good agreement with those obtained by the present JIS method.

According to the Japanese Industrial Standard (JIS) K 0102, we must use a 4-aminoantypyrine (4-AA) batchwise method to determine the quantity of the phenol index after distillation. However, the conventional distillation method must use a large-scale evaporator system, which is time-consuming. In the present work, a miniature evaporator system for phenol index was designed and developed. Also, a sequential injection analysis (SIA) system was proposed for determining the phenol index based on 4-AA spectrophotometry. A calibration curve obtained by the optimized SIA system was linear with a correletion coefficient of 0.998 in the range of 10 mu g L-1 - 10 mg L-1. The limit of detection (3 sigma) was 0.62 mu g L-1, and the limit of quantitation (100) was 2.0 mu g L-1. An SIA peak for phenol was obtained every 75 sec. The performance of the proposed miniature evaporator system was evaluated by distilling standard phenol solutions, followed by the spectrophotometric SIA method. The sample volume for distillation could be reduced to be as small as 35 mL (approximately 1/7 compared with the current JIS procedure) with good recovery. The proposed SIA method coupled with a miniature evaporator system was applicable for determining the phenol index in wastewater.

【目的】当院における過去10年間の脳卒中診療の，内容とその変化について総括した．【方法】1997∼2001年までを前期，2002∼2006年までを後期とし，疾患分布や診療内容について検討した．【結果】総患者数は2,699人（前期1,238・後期1,461），疾患の内訳は虚血性（CI）/脳出血（ICH）/クモ膜下出血（SAH）が前期・後期でそれぞれ59.2%/26.5%/14.3%・62%/24.2%/12.9%であった．治療内容での変化は動脈瘤治療における血管内手術の増加（10%→25%）と，超急性期脳梗塞に対するtPA療法の導入（H16.12∼）であった．予後良好例はCIが，死亡率はSAHが最も多い傾向があった．またICHは機能予後不良例が最も多かった．【結論】当院における脳卒中の治療成績は前後期で大きな改善はなかったが，患者の平均年齢は前後期を比較すると2.4歳高齢化し，平均在院日数は27.2日から23.5日と3.7日短縮した．また，全脳卒中急性期患者の19.6%に外科的治療を要した．

An automated stopped-in-dual-loop flow analysis (SIDL-FA) system is proposed for the determination of vanadium in drinking water. The chemistry is based on the vanadium-catalyzed oxidation reaction of p-anisidine by bromate in the presence of Tiron as an activator to produce a dye (λmax=510nm). A SIDL-FA system basically consists of a selection valve, three pumps (one is for delivering of standard/sample, and others are for reagents), two six-way injection valves, a spectrophotometric detector and a data acquisition device. A 100-μL coiled loop around a heated device is fitted onto each six-way injection valve. A well-mixed solution containing reagents and standard/sample is loaded into the first loop on a six-way valve, and then the same solution is loaded into the second loop on another six-way valve. The solutions are isolated by switching these two six-way valves, so that the catalytic reaction can be promoted. The net waste can be zero in this stage, because all pumps are turned off. Then each resulting solution is dispensed to the detector with suitable time lag. A touchscreen controller is developed to automatically carry out the original SIDL-FA protocol. The proposed SIDL-FA method allows vanadium to be quantified in the range of 0.1–2μgL−1 and is applied to the determination of vanadium in drinking water samples.

特発性正常圧水頭症(iNPH)に対する,圧可変式+重力可変式のハイブリッドなシャントシステムでの治療経験13例を報告した.全例で何らかの症状改善がみられ,術後1年で圧変更を要したのは3例であった.重力可変式システムは,体位にかかわらず生理的な状態に近い脳室内圧が維持できることが,術後の圧測定から裏付けられた.また,調節が不良の場合には圧可変装置が必要であるが,従来の圧可変式のみのシステムと比較すると,圧変更の必要性は非常に少なくできる可能性も示唆された.われわれの用いたハイブリットなシステムは,繊細な調節を必要とするiNPHのシャント治療には,非常に有効である可能性が示された.

We report a case of intracranial hemorrhage due to amphetamine abuse in a young adult. A 34-year-old, confused woman was transferred to our emergency room with right hemiparesis and aphasia. CT at admission demonstrated intracerebral hemorrhage in the left frontal and parietal lobes, associated with subarachnoid hemorrhage. MRA shortly after admission revealed no intracerebral vascular anomaly. Cerebral angiography following admission showed irregularity of the vessel wall in the left anterior and middle cerebral arteries. Later, a toxicology screen test for urine was found to be positive for amphetamines and metamphetamines. These findings suggested that cerebral vasculitis and hypertensive surge induced by amphetamines caused intracranial and subarachnoid hemorrhage. Amphetamine abuse should always be considered as a cause of intracranial hemorrhage in young adults.

The new concept of stopped-in-loop flow analysis (SIL-FA) is proposed, and an SIL-FA method for the catalytic determination of vanadium is demonstrated. In an SIL format, a sample solution merges with reagent(s), and the well-mixed solution is loaded into a loop. The solution in the loop is separated by a six-way switching valve from the main stream. While the reaction proceeds in the stationary loop, the SIL-FA system does not need to establish a baseline continuously. This leads to a reduction in reagent consumption and waste generation compared with traditional flow injection analysis.

A highly sensitive spectrophotometric semi-automated method for ammonium ion determination was proposed using flow injection analysis coupled with an on-line gas diffusion separation/ion exchange concentration technique, which was especially effective for applying to analyses of open sea or fresh seawater. Ammonium ion at the single and/or sub-ppb level in seawater samples was introduced into a gas diffusion system at first, and then subsequently concentrated on a cation exchange resign column. The collected ammonium ion was eluted by 0.5 M hydrochloric acid. After derivatization based on indophenol blue with salicylate, the color development was monitored at 660 nm. The detection limit (3 sigma) for N-NH4+ was 0.5 ppb at 10 minutes concentration and 0.2 ppb at 30 minutes concentration, respectively. The relative standard deviation (n = 5) was found to be 0.68% for 1 ppb of N-NH4+ at 10 minutes concentration. The proposed method could be applicable to the analysis of ammonium ion in seawater samples.

At first, a copper- and/or iron-catalyzed coupling reaction of p-anisidine with N,N-dimethylaniline in the presence of hydrogen peroxide was investigated by batchwise and flow-based methods for the determination of copper and iron in natural water samples. The addition of some ligands (neocuproine for copper and 1,10-phenanthroline for iron) activated the catalytic effects. A flow injection (FI) method using catalytic reactions with a serial flow cell, having two cell compartments, was also applied to the successive assay of labile and inert complexes with humic acid for the characterization of copper and iron complexes. In addition, a sensitive and rapid FI spectrophotometric method was proposed for the determination of cobalt in cobalt alloy, pepperbush leaves and vitamin B-12. It is based on a redox reaction of cobalt(II) with iron (III) in the presence of nitro-PAPS, producing a iron (II)-nitro-PAPS complex. Finally, a sequential injection (SI) method with a lab-on-valve (LOV) format using 5-Br-PSAA was developed for the simultaneous spectrophotometric determination of copper and iron. The computer-controlled SI-LOV system was useful for routine, durable and automated assays of copper and iron in industrial waste water samples. The flow-based analytical techniques with FIA and SI-LOV should be noticed as an environmentally benign technology.

A sequential injection (SI) method in a lab-on-valve (LOV) format for simultaneous spectrophotometric determination of copper and iron has been devised. The detection chemistry is based on the complex formation of 2-(5-bromo-2-pyridylazo)-5-[N-n-propyl-N-(3-sulfopropyl)amino]aniline (5-Br-PSAA) with copper(II) and/or iron(II) at pH 4.6. Copper(II) reacts with 5-Br-PSAA to form the complex which has an absorption maximum at 580 nm but iron(III) does not react. In the presence of a reducing agent only iron(II)-5-Br-PSAA complex is formed and detected at 558 nm. Under the optimum experimental conditions, the determinable ranges are 0.1-2 mg l(-1) for copper and 0.1-5 mg l(-1) for iron, respectively, with a sampling rate of 18 h(-1). The limits of detection are 50 microg l(-1) for copper and 25 microg l(-1) for iron. The relative standard deviations (n=15) are 2% for 0.5 mg l(-1) copper and 1.8% for 0.5 mg l(-1) iron when determined in standard solutions. The recoveries range between 96 and 105% when determining 0.25-2 mg l(-1) of copper and 0.2-5 mg l(-1) of iron in artificial mixtures at copper/iron ratios of 1:10 to 5:1. The proposed SI-LOV method is successfully applied to the simultaneous determination of copper and iron in multi-element standard solution and in industrial wastewater samples.

The reduction reaction of iron(III) with cobalt(II) easily occurs in the presence of 2-(5-nitro-2-pyridylazo)-5-(N-propyl-N-sulfopropylamino) phenol (nitro-PAPS) at pH 3.5, in which iron(II)nitro-PAPS chelate is formed. The chelate has a specific absorption maximum at 790 nm. Cobalt(II) can thus be determined by measuring the absorbance of the complex at the above-mentioned wavelength. The redox reaction is introduced into a two-channel flow system for the determination of cobalt(II). The calibration curve is linear in the concentration range of 2.5 X 10(-7) M similar to 1 X 10(-5) M cobalt(II) with a sample throughput of 70 samples h(-1). The detection limit (S/N = 3) is 1 X 10(-7) M and the relative standard deviation for 10 determinations is 0.3% for 2.5 X 10(-6) M cobalt(II). The proposed flow-injection spectrophotometric method is applied to analyses of cobalt(II) in cobalt alloy (NIST SMR 862; High Temperature Alloy L 605) and pepperbush (NIES standard, No. 1) and to the indirect determination of vitamin B-12 in pharmaceuticals.

A flow injection-catalytic spectrophotometric method using a serial flow cell was proposed for the successive determination of trace amounts of copper and iron. This method is based on the oxidation coupling of p-anisidine with N,N-dimethylaniline in the presence of hydrogen peroxide to form a dye, which has an absorption maximum at 740 nm. In this indicator reaction, ligands such as 1,10-phenanthroline (phen) and diphosphate were achieved to improve the sensitivity and selectivity. Under the optimal experimental conditions, the determinable ranges were 0.05 - 5 ppb for copper and 0.5 - 100 ppb for iron, respectively. The RSDs (n = 10) were 0.78% for 0.5 ppb copper(II) and 0.5% for 200 ppb iron(III). The sample throughput was 30 h(-1). The present flow-injection method was applied to the determination of copper and iron in standard river water, tap water, and other natural water samples, and also to the analysis of labile and inert complexes in synthesized samples containing humic acid with copper(II) or iron(III).

A kinetic–catalytic spectrophotometric method is proposed for the successive determination of nanogram levels of copper and iron, which is based on their catalytic effects on the oxidative coupling of p-anisidine with N,N-dimethylaniline (DMA) to form a colored compound (λmax=740 nm) in the presence of hydrogen peroxide at pH 3.2. 2,9-Dimethyl-1,10-phenanthroline (neocuproine) acted as an activator for the copper catalysis, and 1,10-phenanthroline (phen) acted as an activator for the iron catalysis. The selectivity was improved in the presence of diphosphate as a masking agent. The determinable ranges were 0.16–10 ppb for copper and 1–100 ppb for iron, respectively. The relative standard deviations of copper and iron were 1.1 and 0.97% for five determinations of 10 ppb copper and 40 ppb iron. The method was successfully applied to the analyses of copper and iron in tap, well, river and pond waters.

The cells from plasmacytomas induced in BALB/c and susceptible BALB/c congenic strains of mice by pristane (2,6,10,14-tetramethylpentadecane) (1) have non-random chromosomal translocations, rcpt (6;15) or rcpt(12;15) (2, 3). To date, the karyotypes of 27 early transfer generation or primary pristine-induced plasmacytomas have determined, and 26 of the tumors have either rcpt(6; 15) or rcpt(12;15) (3-5). The transiocation breakpoints on these chromosomes occur in the same chromosome bands in all the plasmacytomas studied to date. The breakpoints in chromosomes 12 and 15 have been studied by cloning and DNA sequencing. The breakpoint in chromosome 12 occurs in the immunoglobulin heavy chain gene (IgH) ~ complex, frequently, but not always, in the switch region of the alpha heavy chain gene (6-11). In chromosome 15 the breakpoint occurs close to or within the c-myc oncogene (8-12). The breakpoints in chromosome 6 have not yet been determined. Expression of c-myc RNA, often in relatively large amounts, characterized most of the plasmacytomas (11, 13, 14), and rearrangement of c-myc DNA was frequently seen. The development of plasma cell tumors in BALB/c mice injected with pristane alone requires a minimum of 120 d, but usually more than 200 d (15). In contrast, plasmacytomagenesis can be greatly accelerated (5, 16) by infecting pristane-injected mice with Abelson virus. This murine acute leukemia virus contains two retroviral elements, a replication competent Moloney leukemia virus helper and a defective component, A-MuLV (17). The defective element has lost part or all of the viral gag, pol, and env genes and has acquired sequences from the mouse c-abl gene (for review see reference 18). Adult mice of only a This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan, and by grants from the National Cancer Institute, Dept. of Health and Human Services, and The Swedish Cancer Society. 1 Abbreviations used in this paper: ABPC, plasmacytomas induced by Abelson virus plus pristane; A-MuLV, Abelson murine leukemia virus; IgH, immunoglobulin heavy chain; i.p., intraperitoneal; LTR, long terminal repeats of retroviruses; rcpt, reciprocal chromosome transiocation.

A kinetic-catalytic spectrophotometric flow-injection method was developed for the rapid and sensitive determination of trace amounts of copper(II). The method is based on the catalytic effect of copper(II) on the redox reaction of cysteine with iron(III). Iron(II) produced by the catalytic reaction reacts with 2,4,6-tris(2-pyridyl)-1,3,5-triazine (TPTZ) to form the iron(II)-TPTZ complex (λmax = 593 nm). By measuring an absorbance of the complex, one could determine 0.05–8 ppb copper(II) with the relative standard deviations (n = 10) of 1.6%, 1.3%, and 0.8% for 0.5 ppb, 1 ppb, and 2 ppb copper(II), respectively. The limit of detection (S/N = 3) was 0.005 ppb. The sample throughput was 30 h−1. The proposed method was successfully applied to the determination of copper in natural water and serum samples.

Subcellular localization and transactivation of human hepatitis B virus X protein (HBx), a plausible causative factor for hepatocellular carcinogenesis, were studied in transiently transfected cells. The transactivation was detected not only by the cis-element driven chloramphenicol acetyltransferase (CAT) assay but also by immunostaining of CAT protein cotransfected into human hepatoma cell line HepG2. Scanning fluorescence microscopy showed the majority of immunological signals of HBx to be at the perinuclear region of transfected cytoplasm. HBx was also clearly detectable in the nucleus, though less intensely expressed. This was confirmed by Western analysis and coimmunoprecipitation of HBx with transcription factor IIB (TFIIB) in subcellular fractionations. The percentage of HBx-positive cells coincided with that of CAT-positive cells, and confocal laser microscopy revealed the coexistence of CAT signals in GFP-HBx positive cells. The SV40 large T antigen nuclear localization signal (NLS) appended HBx, regardless of whether NLS was added to the N- or C-terminus, transactivated all the examined X-responsive elements (XRE) similarly as did wild-type HBx. Similar results were obtained in p53 negative Saos-2 cells. The detected nuclear HBx may be involved in modulating the transcription at the promoter level whereas the HBx in cytoplasm may be working through signal transduction pathways.

ADAM is a recently discovered gene family that encodes proteins with a disintegrin and metalloproteinase. ADAMTS-1 is a gene encoding a new member protein of the ADAM family with the thrombospondin (TSP) type I motif, the expression of which is associated with inflammatory processes. In the present study, we have characterized the exon/intron organization of the mouse ADAMTS-1 gene. The ADAMTS-1 gene is composed of nine exons, all of which are present within the 9.2-kb genomic region. Among the nine exons, exons 1, 5, and 6 encode a proprotein domain, a disintegrin-like domain, and a TSP type I motif, respectively, of the ADAMTS-1 protein, suggesting that there is a correlation between exon/intron organization and functional domains. In addition, the exon/ intron organization of the ADAMTS-1 gene is very different from that of the metalloproteinase-like/disintegrin-like/ cysteine-rich protein gene (MDC) (ADAM11), suggesting that the genomic structure of ADAM family genes is not necessarily conserved. Furthermore, fluorescence in situ hybridization revealed that the ADAMTS-1 gene is located in region C3-C5 of chromosome 16, to which none of the previously identified ADAM genes have been mapped.

Human hepatitis B virus (HBV) X protein, HBx, transactivates virus and host genes through a wide variety of cis-elements. Expression of HBx is controlled by HBV enhancer 1 (Enh1). Both Enh1 and the core sequence of Enh1, which consists of an AP-1 related site (cFAP1) and a C stretch, respond to HBx and a phorbol ester (TPA). Biochemical pathways of the responses to HBx and TPA are still controversial. We therefore asked whether HBx and TPA stimulate Enh1 core activity through a common process. Protein kinase C (PKC) inhibitors, H-7 and staurosporin, did not inhibit HBx transactivation at concentrations sufficient to abolish the TPA effects in HepG2 cells. Although HBx transactivation synergized independently with TPA or a phosphoprotein phosphatase inhibitor, okadaic acid (OA), the PKC inhibitors eliminated only the TPA contribution. HBx transactivation required both the cFAP1 and the C stretch of the Enh1 core region; however, mutations in either or both of the two cis-elements demonstrated that TPA augmentation required only cFAP1. These results imply that HBx transactivation operates through a mechanism distinct from the PKC and OA activation pathways.

The DCPC 21 plasmocytoma lacks any of the MPC‐associated chromosomal translocations. However, the c‐myc gene has been transposed to the IgH locus on chromorome 12 by an Ig switch‐region‐mediated recombination mechanism. DNA sequencing analysis, further, revealed that this recombination is consistent with an insertion of the IgH enhancer (Eμ)‐Sμ sequences, 2341 bp in length, into the c‐myc 5′‐flanking region, resulting in 5′: c‐myc 5′‐flanking‐Eμ‐Sμ‐c‐ myc 5′‐flanking‐ c‐myc exon‐1 : 3′ segment. In situ molecular hybridization of DCPC 2I metaphase chromosome spreads using a Pvt‐1 probe demonstrated that Pvt‐1 has also moved to the FI sub‐band region of chromosome I2 where the IgH genes are located. These results indicate that the c‐myc gene has been inserted into the IgH locus together with the Pvt‐I, regardless of whether plasmocytoma has cytogenetically identifiable translocations. The possible interaction between c‐myc activation and Pvt‐I in the development of MPCs is discussed.