Multilayer coated grazing incidence condenser for large numerical aperture objective at wavelength of 4.5 nm.

A grazing incidence condenser is developed for objectives with large numerical aperture working in Carbon-window wavelength region (λ=4.4-5.0 nm) with the use of a point light source. The condenser is composed of four pieces of toroidal mirrors and a piece of the mirror was fabricated to evaluate the performance of the mirror. The radii of the toroidal mirror are determined by ray-trace calculation, and each radius of the mirror substrate and the roughness of the polished surface were evaluated to satisfy the designed parameter. A Co/C reflection multilayer is also designed to reflect soft x-ray light at 4.5 nm wavelength, and the reflection multilayer was deposited on the mirror surface. Measured reflectance of the toroidal mirror with the reflection multilayer is higher than 0.32 at 4.5 nm wavelength.

Minimal deviation endometrioid adenocarcinoma of the endometrium and its MRI findings.

Minimal deviation endometrioid adenocarcinoma (MDA-E) of the endometrium is a rare pathological entity, and its radiological features are rarely documented. A 73-year-old Japanese woman was referred to the authors when an endometrial biopsy revealed moderately differentiated endometrioid adenocarcinoma. Preoperative radiological examinations, including ultrasound, computed tomography (CT), and magnetic resonance imaging (MRI) showed no evidence of cancer nests. In the hysterectomy specimen, mildly atypical glands were scattered throughout the entire depth of the myometrium, without stromal desmoplastic reaction, and a tiny focus of typical, ruptured, endometrioid adenocarcinoma glands was found in the atrophic endometrium. MRI had not been able to identify this unusual, scattered, myometrial invasion. It should be kept in mind that in cases showing Stage IA endometrial carcinoma without endometrial thickening on MRI, this rare form of invasion may be present.

A wavelength modulation system for highly sensitive absorption spectroscopy.

We developed a newly designed wavelength modulation (WM) system for highly sensitive absorption spectroscopy. In our system, the WM is realized by yawing an output mirror in a monochromator. In order to control an amplitude Δλ of the WM in a wide range, we employed a forced vibration of a permanent magnet driven by a magnetic field of a solenoid. Our system has an advantage of that the WM amplitude Δλ can be adjusted in extensively wide range from 0.08 nm to 11 nm only by tuning a driving frequency of the applying current to the solenoid, because we utilize a resonance phenomenon of the forced vibration for adjustment of the WM amplitude. By using our system, we measured WM absorption spectra of a Cu(2)O thin film and found clearly spectral structures for weak 2-4P excitonic resonances in the WM absorption spectra.

Chitin synthase 1 (Chs1) gene sequences of Microsporum equinum and Trichophyton equinum.

Chitin synthase 1 (Chs1) genes from Microsporum equinum and Trichophyton equinum were compared with those of the other dermatophytes. The Chs1 nucleotide sequences of these dermatophytes from horses showed more than 80% similarity to those of Arthroderma benhamiae, A. fulvum, A. grubyi, A. gypseum, A. incruvatum, A. otae, A. simii, A. vanbreuseghemii, Epidermophyton floccosum, T. mentagrophytes var. interdigitale (T. interdigitale), T. rubrum and T. violaceum. Especially high degree of nucleotide sequence similarity of more than 99% was noted between the Chs1 gene fragments of M. equinum and A. otae, and those of T. equinum, T. interdigitale and A. vanbreuseghemii, respectively. The phylogenetic analysis of their sequences revealed that M. equinum was genetically very close to A. otae and T. equinum to A. vanbreuseghemii. A molecular analysis of Chs1 genes will provide useful information for the genetic relatedness of M. equinum and T. equinum and confirm the value of DNA sequencing in identification of these two dermatophytes.

Use of a temporary caval filter in a young man with pulmonary embolism to prevent migration of massive caval thrombus during an attempt of caval thrombolysis.

A 33 year old male with no known risk factors for hypercoagulability developed a massive thrombi in the inferior vena cava (IVC). The patient had a history of both pulmonary embolism and embolism related syncope. The thrombus which extended proximally to the level of the renal vein and distally to the left superficial femoral vein did not respond to anticoagulant therapy or thrombolysis. Thirteen days after admission, we decided to use a temporary caval filter to provide protection from migration of the thrombus while attempting invasive thrombolytic therapy, which was performed using a tissue type plasminogen activator through a coaxial catheter of the temporary filter. This resulted in a marked decrease in the size of the thrombus, and multiple thrombi were found to be trapped in the temporary filter. Although the temporary caval filter was effective in capturing emboli, resulting in a decrease in the thrombus size, the thrombus was not completely dissolved within two weeks, which is the maximal implantation time. A permanent filter was eventually used to prevent pulmonary embolism, which could arise from the remaining thrombus. We have found placement of a temporary caval filter to be a safe and effective adjunct, in select cases, when attempting thrombolysis of massive thrombi in the IVC. Since we inserted the temporary filter 13 days after admission, use of a temporary filter during thrombolysis may have been more effective if conducted earlier in our patient's clinical course.

Polyangiitis overlap syndrome with eosinophilia associated with an elevated serum level of major basic protein.

Polyangiitis overlap syndrome is a new disease entity and the reported cases in the literature are still limited. We describe a female patient presenting with finger ulcers, skin eruptions, pleural effusion, interstitial pneumonia and eosinophilia. Skin biopsy showed systemic small-sized angiitis and thrombosis. She was diagnosed as having polyangiitis overlap syndrome and was successfully then treated with corticosteroid. It is also of interest that the disease activity was correlated with the number of eosinophils in peripheral blood. The measurement of the serum level of major basic protein released from eosinophils functioning as a coagulant indicated the possible association of eosinophilia with thrombosis and polyangiitis.

[Combination of 5'-DFUR and MMC for recurrent rectal cancer with pulmonary metastasis].

We report a 68-year-old man with rectal cancer and recurrent pulmonary metastasis treated with concomitant 5'-DFUR + MMC, which resulted in an extreme reduction of the lesion. The initial pulmonary metastases were treated by pneumonectomy, but 32 months later the patient again showed a pulmonary metastasis. Thus, from July 1997, he was treated with 5'-DFUR (800 mg/day) + MMC (4 mg/2 weeks). After 3 months of therapy, the chest CT examination showed an extreme reduction of the pulmonary lesion, and at 5 months the lesion was even smaller. Presently, after 7 months of therapy, the lesion remains stable. His CEA level is 1.0-2.6 ng/ml. After 4 months of treatment he had mild anorexia, which was alleviated by reducing the 5'-DFUR to 600 mg/day. No other adverse reaction was observed; the therapy was safely conducted on an outpatient basis.

Effect of a novel non-steroidal anti-inflammatory drug (M-5011) on cytokine levels in rats with monosodium urate crystal- induced pleurisy.

We evaluated the effects of a new non-steroidal anti-inflammatory drug (NSAID), d-2-[4-(3-methyl-2-thienyl)phenyl]propionic acid (M-5011), and indomethacin on the production of arachidonate metabolites and pro-inflammatory cytokines in male Sprague-Dawley rats with monosodium urate crystal (MSU)-induced pleurisy. Levels of tumor necrosis factor (TNF), interleukin (IL)-1 and IL-6 in the pleural exudate were determined by biological assays, while prostaglandin E2 (PGE2), leukotriene B4 (LTB4) and cytokine-induced chemoattractant-1 (CINC-1) levels were quantified by enzyme immunoassays. Orally administered M-5011 (5 mg/kg) decreased the pleural exudate volume at 3 and 4 hr after MSU injection. Indomethacin (10 mg/kg) decreased the volume at 3-5 hr. These drugs reduced the number of leukocytes in the pleural cavity at 6 hr. Both NSAIDs also reduced the content of PGE2 in the exudate without affecting LTB4 levels. Increased productions of both IL-6 and CINC-1 in the exudate were reduced by pretreatment with M-5011 or indomethacin, and TNF levels in the exudate were increased by pretreatment of these drugs. Thus, M-5011 inhibits the production of both IL-6 and CINC-1 at lower doses than those of indomethacin, and the inhibitory effect of M-5011 on CINC-1, but not IL-6, may partly contribute to the inhibition of leukocyte infiltration in rats with MSU-induced pleurisy.

An aggressive desmoid tumor in a patient with familial adenomatous polyposis: immunohistochemical findings.

A case of an aggressive desmoid tumor in a patient with familial adenomatous polyposis is described. The lesion rapidlyenlarged with compression of adjacent structures including the ureter and small bowel, and the patient died because of small bowel perforation and hydronephrosis 3 years after detection of small desmoid tumors at the time of a prophylactic coloproctectomy for a colon carcinoma. Immunohistochemically, proliferating cell nuclear antigen (PCNA), p21WAF1/CIP1 and cathepsin D indices, but not the bcl-2 index, which were defined as the numbers of immunoreactive tumor cells per 1000 tumor cells, increased in line with tumor progression. The tumor did not show staining for collagen IV, but was characterized by intense staining for basic fibroblast growth factor (bFGF). Accordingly, tumor aggression was related to increases in both cell proliferation and protease activity, as well as an enhanced expression of bFGF. In addition, the desmoid tumor showed deregulation between PCNA and p21WAF1/CIP1 because the normal inverse relation between these two was not apparent.

Human papillomavirus oncoprotein E6 binds to the C-terminal region of human minichromosome maintenance 7 protein.

Oncoprotein E6 of the human papillomavirus (HPV) associated with cervical cancer (HPV-16 and -18) degrades tumor suppressor protein p53, but seems to have p53-independent transforming functions. We searched for other cellular targets for the N-terminal region of HPV-16 E6 using a yeast two-hybrid system. The E6 was found to bind to the C-terminal region of a human minichromosome maintenance 7 (hMCM7) protein, which is a component of replication licensing factors. The full-length hMCM7 translated in vitro was capable of binding to bacterially expressed E6. In yeast cells the E6s of the cancer-associated HPVs (HPV-16, -18, and -58) bound to hMCM7 more strongly than those of the HPVs associated with a benign tumor (HPV-6 and -11). Binding of E6 with hMCM7 may cause chromosomal abnormalities found in the human cells expressing E6s of oncogenic HPVs.

Inhibitory action of indomethacin on neutrophil infiltration in monosodium urate-induced pleurisy in rats.

The effects of indomethacin on the production of cytokines at inflammatory sites were investigated in the monosodium urate (MSU) pleurisy model characterized by both cellular influx and edema. Indomethacin (10 mg/kg) orally administered 0.5 hr prior to MSU injection into the pleural cavity significantly inhibited MSU-induced neutrophil accumulation in the cavity. In addition, the drug slightly enhanced the level of MSU-induced tumor necrosis factor production without affecting interleukin-1 production. Furthermore, indomethacin inhibited both the levels of MSU-induced rat cytokine-induced neutrophil chemoattractant (CINC/gro) and interleukin-6 (IL-6) production by 78.3% at 3 hr and 45.8% at 4 hr post-injection, respectively. Although intrapleural injection of CINC/gro induced neutrophil infiltration in a dose-dependent manner, IL-6 did not affect the action of CINC/gro on neutrophil influx. These findings suggest that the inhibitory action of indomethacin on neutrophil infiltration is, at least, partly mediated by a decrease in the MSU-induced CINC/gro content in this model.

[Usefulness of intracardiac echocardiography for guidance of transseptal puncture procedure].

The clinical usefulness of intracardiac echocardiography (ICE) for the guidance of transseptal puncture procedure during percutaneous left heart bypass support (PLHBS) and percutaneous transvenous mitral commissurotomy (PTMC) was investigated to replace intraoperative transesophageal echocardiography monitoring which requires mild sedation and causes patient discomfort. The ICE procedure was assessed in 3 patients with PLHBS and 18 with PTMC using a 10 MHz rotating 8 French probe system especially developed for the purpose. Transseptal puncture procedure was observed by intraoperative ICE monitoring in the right atrium. The ICE images showed the transseptal puncture (Brokenbrough) needle as a point casting an acoustic shadow. By moving the ICE probe up and down, the excursion of the needle and its approach to the septal wall could be clearly observed. If the puncture needle is forced into the intra-atrial septum, the septal wall was clearly observed to protrude into the left atrium (tent-formation). In a tent-formation the puncture site could be determined by ICE alone. Intracardiac echocardiography guidance was useful and may improve the safety and reliability of the transseptal puncture procedure in PLHBS and PTMC.

Demonstration of sugar moiety on the surface of hepatitis C virions recovered from the circulation of infected humans.

The amino acid sequence for the envelope protein(s) predicted from the nucleotide sequence of the E and E2/NS1 regions of the hepatitis C virus (HCV) genome is enriched with an N-linked glycosylation site motif, Asn-X-Thr/Ser, suggesting oligosaccharide moieties are present on the virion surface. We attempted to characterize the sugar moiety on the surface of HCV virions recovered from sera of infected humans to assess the natural properties of the virus. Six kinds of lectins were used to bind HCV virions in affinity column chromatographies: RCAI, WGA, Con A, AAL, LCA, and PNA. Lectin-bound virions were identified by detecting HCV RNA in eluted chromatography fractions with a polymerase chain reaction (PCR) method. Our results showed that HCV was similar to hepatitis B virus (HBV) in characteristics of binding to lectins: HCV showed a strong binding to RCAI and WGA, weak binding to Con A, and no detectable binding to AAL, LCA, or PNA. Treatment of the HCV virion preparation with an enzyme, glycopeptidase A, or a detergent, NP-40, resulted in a significant decrease in the ability to bind these lectins. Our results suggest that asparagine-linked sugar chains are present on the surface of native virions of HCV, very similar to those for HBV.

[Effects of sodium hyaluronate on the nociceptive response of rats with experimentally induced arthritis].

Antinociceptive effects of sodium hyaluronate (Na-HA) were studied on the basis of improvement in the graded abnormal gait elicited by arthritis induced by intra-articular administration of monosodium urate crystal (MSU) to rats. One hour before MSU injection, intra-articular administration of a 1.0% solution of Na-HA with different molecular weights, ranging from 4.70 x 10(5) to 2.02 x 10(6) (HA-200), improved the score of abnormal gait in a molecular weight-dependent manner in the experimental arthritis model. Similarly, administrations of HA-200 at concentrations ranging from 0.1 to 1.0% prior to MSU treatment resulted in improvement of the score in abnormal gait in a dose-dependent manner. To elucidate the antinociceptive mechanisms of Na-HA, effects of pretreatment with Na-HA (1.0%) of different molecular weights on prostaglandin E2 (PGE2) and bradykinin (BK) releases in synovial fluid 3 hr after MSU injection were studied. Increases in PGE2 and BK concentration in the synovial fluid were depressed in a molecular weight-dependent manner by Na-HA (1.0%) pretreatment. These results indicate that Na-HA attenuates the nociceptive responses inflicted by the MSU-induced arthritis. Such an antinociceptive effect may be due to the inhibition of PGE2 and BK synthesis in the synovial joint of rats.

High-performance liquid chromatographic measurement of tryptophan in blood, tissues, urine, and foodstuffs with electrochemical and fluorometric detections.

A rapid and convenient measurement of tryptophan in whole blood, serum, liver, brain, urine, and alkaline hydrolysates of proteins and foodstuff was done by high-performance liquid chromatography. The sample preparation was simply homogenized or mixed in a 5% trichloroacetic acid solution and a sample of the supernatant was injected onto a column after filtration with a 0.45-micron filter. The method used a Chemcosorb 5-ODS-H column (particle size, 5 microns, 150 x 4.6 mm i.d.) eluted with 20 mM potassium dihydrogen phosphate (pH adjusted to 3.7 by the addition of phosphoric acid) containing 1 g/l of sodium heptane sulfonate and 3 mg/l EDTA.2Na-acetonitrile (93:7, v/v) at a flow rate of 1.5 ml/min. The tryptophan contents in whole blood, serum, liver, and brain were electrochemically estimated at +1000 mV vs. Ag/AgCl, the detection limit being 0.2 pmol (40.84 pg) at a signal-to-noise ratio of 5:1. The tryptophan contents in urine, proteins, and foodstuff were fluorometrically estimated with an excitation wavelength of 280 nm and with an emission wavelength of 340 nm, the detection limit being 20 pmol (4.08 ng) at a signal-to-noise ratio of 5:1. Tryptophan was eluted at about 10.5 min. The total analysis time was about 12 min.

Identification of mutations that occurred on the genome of Japanese encephalitis virus during the attenuation process.

The total nucleotide sequences of the genomes of two Japanese encephalitis virus (JEV) strains, the attenuated vaccine strain SA14-14-2 and its parental virulent strain SA14, were determined by using the molecular cloning technique. The sequence analysis revealed that both virion RNA molecules were 10,976 nucleotides long with 95 and 585 flanking bases at the 5' and 3' untranslated sequences, respectively. A single, long open reading frame spanning 10,296 nucleotides was observed to encode a polyprotein of 3432 amino acid residues. When these sequences were compared with each other, 57 nucleotide substitutions were found to be scattered all over the genome. Of these, 24 resulted in amino acid changes within viral proteins. Structural proteins C and E contain one and eight amino acid changes, respectively. Of the nonstructural proteins, NS1 contains three, NS2a two, NS2b two, NS3 four, NS4a one, NS4b one, and NS5 two amino acid substitutions. The 5'- and 3'-terminal untranslated regions contain one- and two- point mutations, respectively. These data and comparative studies with other JEV strain genomes provide a molecular basis for investigating attenuation mechanisms of JEV.

Combined hepatitis B immune globulin and vaccine for postexposure prophylaxis of accidental hepatitis B virus infection in hemodialysis staff members: comparison with immune globulin without vaccine in historical controls.

Twenty-three staff members serving in a hemodialysis unit were exposed accidentally to needlestick contaminated with blood containing hepatitis B surface antigen and hepatitis B e antigen, as well as high levels of DNA polymerase activity (greater than 100 cpm). They received hepatitis B vaccine (20 micrograms) simultaneously with hepatitis B immune globulin (5 ml, 200 IU per ml) within 48 hr after the exposure, and the vaccination was repeated at 1 and 3 months. The protective efficacy was compared with that in a past study in the same unit in which 33 members were given hepatitis B immune globulin alone within 48 hr after the exposure to blood with similarly high levels of DNA polymerase activity. No differences were noted in age or sex between the staff members who were vaccinated and those who were not, nor were there any differences between their inocula in the titers of hepatitis B virus markers. During 12 months after the accident, only one (4%) of the 23 vaccinated members contracted hepatitis B virus infection, at a frequency significantly lower than 11 (33%) of the 33 members who did not receive vaccine (p less than 0.02). These results indicate that hepatitis B vaccine, when given in combination with hepatitis B immune globulin, is efficacious for postexposure immunoprophylaxis of accidental infection.

Antibodies to translation products of the pre-S1 and pre-S2 regions of the envelope gene of hepatitis B virus in fulminant hepatitis B.

Sera from 11 patients with fulminant hepatitis B were tested for antibodies to translation products of the pre-S1 and pre-S2 regions of hepatitis B virus of IgM, IgA and IgG classes, as well as of IgA1, IgA2 and SIgA, with solid-phase enzyme immunoassays using native viral polypeptides. Antibodies to pre-S1 region product of IgM and/or IgA class were detected invariably in six patients who still had detectable hepatitis B surface antigen in serum at the time of clinical presentation. The remaining five patients who had lost HBsAg at presentation had antibodies to pre-S region products of various immunoglobulin classes in higher titers. The five patients with fulminant hepatitis without HBsAg had higher levels of IgA antibodies to pre-S region products than the seven patients with nonfulminant acute hepatitis B who had lost HBsAg: IgA antibody to pre-S1 region product (75.6 +/- 63.8 vs. 2.9 +/- 3.2, p less than 0.01) and IgA antibody to pre-S2 region product (28.9 +/- 25.3 vs. 4.2 +/- 6.9, p less than 0.01). IgA antibodies to pre-S1 and pre-S2 region products were invariably polymeric in fulminant hepatitis B. These findings are compatible with the hypothesis that a heightened humoral antibody response to pre-S1 and pre-S2 region products occurs early during the course of fulminant hepatitis B, participating in severe hepatic injury and early clearance of virus characteristic of this disease.

Small and large forms of hepatitis B e antigen in the serum: determination by two-site sandwich radioimmunoassay with monoclonal antibodies.

Serum samples containing hepatitis B e antigen (HBeAg) were subjected to electrophoresis in agarose, and fast-migrating 'small' HBeAg and slow-migrating 'large' IgG-associated HBeAg were pooled separately. HBeAg of each category was determined by a two-site radioimmunoassay that sandwiched HBeAg between monoclonal antibody (anti-HBe) against one epitope of HBeAg, fixed on a solid support, and anti-HBe against another epitope, labelled with radioiodine. Eighteen sera in which small HBeAg dominated revealed activities of hepatitis B surface antigen-associated DNA polymerase significantly higher than 14 sera in which large HBeAg dominated (logarithm of ct/min, mean +/- s.e. 3.36 +/- 0.08 versus 2.14 +/- 0.11, P less than 0.01). Shift from small HBeAg to large HBeAg was observed, along with the disappearance of DNA polymerase, in the serum from two carriers who seroconverted to anti-HBe.