Inscription of silicon waveguides using picosecond pulses.

Direct writing of single-mode waveguides into crystalline silicon using ps laser pulses is presented. The embedded structures were fabricated by moving the focal position along the beam axis with the help of a long distance microscope objective. In situ monitoring during inscription was performed to analyze the processing dynamics. The waveguide generation is based on pronounced multi-pulse interaction at moderate pulse energies around 100 nJ. All samples were characterized in terms of mode field distribution and damping losses. Calculations indicate an induced refractive index change in the range of 10-3 to 10-2. Moreover, a Y-splitter was realized to demonstrate the potential of this process.

Antiviral activity of hypothiocyanite produced by lactoperoxidase against influenza A and B viruses and mode of its antiviral action.

Hypothiocyanite (OSCN-) is a natural component of human saliva and is produced by the lactoperoxidase (LPO)/thiocyanate/hydrogen peroxide (H2O2) system. OSCN- has been previously shown to exhibit antiviral activity against influenza viruses (IFV) A/H1N1/2009 and A/H1N2/2009 in vitro as well as antimicrobial and antifungal activities. We elucidated the antiviral activity of OSCN- against both IFV types A and B and the mode of its antiviral action. OSCN- was produced constantly at 900 ± 200 µmol/l in Na3PO4 buffer solution containing NaSCN and LPO in the presence of H2O2 as an original OSCN- solution. In a plaque reduction assay, IFV A/PR/8/34 (H1N1), A/Fukushima/13/43 (H3N2), B/Singapore/222/97, and B/Fukushima/15/93 were exposed to various concentrations of OSCN- for 0 to 30 min before adsorption to MDCK cells, and plaque formation was examined. OSCN- exhibited significant similar antiviral activities against all four viruses without cytotoxicity, and the EC50 values for them were from 57 ± 16 to 148 ± 27 µmol/l regardless of the exposure times. The exposure of MDCK cells to OSCN- before viral adsorption did not affect its anti-IFV activity (EC50: more than 450 µmol/l), but the exposure after viral adsorption affected it moderately (EC50: 380 ± 40 µmol/l). Moreover, the exposure of virus particles to OSCN- at 450 µmol/l did not affect the hemagglutinin activity of IFV in hemagglutination inhibition assay. These results suggest that the attachment of OSCN- to the viral envelope critically contributes to the mode of antiviral action of OSCN- without interfering with viral adsorption. Keywords: hypothiocyanite; influenza virus type A; influenza virus type B; lactoperoxidase; antiviral activity.

Corticosteroids plus long-acting ß2-agonists prevent double-stranded RNA-induced upregulation of B7-H1 on airway epithelium.

BACKGROUND: Airway viral infections provoke exacerbations of asthma and chronic obstructive pulmonary disease. B7-H1 is a costimulatory molecule that is implicated in an escape mechanism of viruses from host immune systems. This escape may be associated with the persistence of viral infection and lead to exacerbation of underlying diseases. We have shown that an analog of viral double-stranded RNA, polyinosinic-polycytidylic acid (poly IC), upregulated the expression of B7-H1 on airway epithelial cells, an effect which was corticosteroid-resistant. We investigated the effects of corticosteroids plus long-acting ß(2)-agonists (LABAs; fluticasone/salmeterol or budesonide/formoterol) on the expression of B7-H1. METHODS: BEAS-2B cells and primary airway epithelial cells were stimulated with poly IC or respiratory syncytial virus. The expression of B7-H1 was assessed by flow cytometry. RESULTS: Poly IC upregulated the expression of B7-H1, which was suppressed by high-concentration corticosteroids but not by LABAs. The upregulation was suppressed by very low-concentration corticosteroids when used in combination with LABAs. Their combination also suppressed the virus-induced upregulation of B7-H1. Poly IC stimulation induced the nuclear translocation of nuclear factor ĸB (NF-ĸB). Inhibitors of NF-ĸB activation prevented the poly IC-induced upregulation of B7-H1. Low-concentration corticosteroids in combination with LABAs enhanced the de novo induction of IĸBα, the endogenous inhibitor of NF-ĸB activation. CONCLUSIONS: Fluticasone/salmeterol or budesonide/formoterol attenuate the virus-associated upregulation of B7-H1 on airway epithelial cells via suppression of NF-ĸB activation.

Interleukin 12 is a primary cytokine responding to influenza virus infection in the respiratory tract of mice.

We have reported previously that an increase in interleukin 12 (IL-12) production in the lungs of mice infected with Influenza A virus or an intranasal (i.n.) administration of IL-12 to the infected mice alleviated pneumonia (Tsurita et al., J. Pharmacol. Exp. Therapeut. 298, 362-368, 2001). In this study, we found that in the bronchoalveolar lavage fluids (BALF) obtained from mice infected i.n. with Influenza A virus IL-12 was elevated on day 1 post infection (p.i.) and was followed by tumor necrosis factor alfa (TNF-alfa), IL-18, and interferons alfa, beta, and gama (IFN-alfa, -beta, and -gama) on day 2 p.i. Histochemical analyses of the infected lungs on day 1 p.i. showed the presence of IL-12 and IL-12 mRNA in mononuclear and macrophage-like cells and co-localization of macrophages with viral antigen, while other cytokines were absent. Thus, IL-12 was produced by macrophages infiltrating the infected epithelium as the first response cytokine and its production at the site of infection may direct an early immune defense to alleviate the severity of infection.

Establishment of CHO cell lines expressing four N-methyl-D-aspartate receptor subtypes and characterization of a novel antagonist PPDC.

To develop an assay system that allows the N-methyl-D-aspartate (NMDA) receptor subtype-selective antagonistic potency of drugs, we have established Chinese hamster ovary cell lines expressing the four NMDA receptor subtypes (GluRepsilon1/zeta1-GluRepsilon4/zeta1) heat-indelibly. Using these clonal cells, we found that a novel antagonist, (1S,2R)-1-phenyl-2[(S)-1-aminopropyl]-N,N-diethylcyclopropanecarboxamide, was less selective for the GluRepsilon1/zeta1: the IC(50) values for the GluRepsilon1/zeta1-GluRepsilon4/zeta1 were 41.7, 13.3, 12.6 and 11.5 microM, respectively, while two well-known antagonists, DL-2-amino-5-phosphonovaleric acid and ifenprodil, showed the known potency and selectivity for each subtype. Thus, the established clonal cells are of use in characterizing the pharmacological properties of drugs that act on NMDA receptors.

Fabricating micro-Bragg reflectors in 3-D photorefractive waveguides.

Bragg reflectors as small as optical waveguides can be used at the intersections of waveguides as spectral filters, beam splitters, or beam combiners. We present the concept of fabricating small Bragg reflectors in photorefractive waveguides and show the spectral selectivity of these micro-Bragg reflectors fabricated in a lithium niobate crystal. We also show that we can fabricate waveguide structures simultaneously with the micro-Bragg reflector, by using a crystal whose c-axis is directed 45 degree off the optical axis of the fabrication beam. The reflectivity of the micro-Bragg reflector was smaller than 0.01 and was not large enough for the immediate use.

Inducible expression of N-methyl-D-aspartate (NMDA) receptor channels from cloned cDNAs in CHO cells.

To develop a drug screening system, we introduced expression vectors carrying the mouse N-methyl-D-aspartate (NMDA) receptor channel epsilon1 and zeta1 subunit cDNAs under the promoter of the Drosophila heat shock protein hsp70 into Chinese hamster ovary (CHO) cells. We selected clonal cell lines by means of RNA blot hybridization and fura-2 fluorometry. One of these cell lines, ZE1-1, optimally expressed the epsilon1 and zeta1 subunit mRNAs when induced by an incubation at 43 degrees C for 2 h. Heated ZE1-1 cells exhibited the NMDA-induced intracellular Ca2+ elevation, whereas unheated they showed no such response. NMDA and L-glutamate, but not alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and kainate, induced an increase in the intracellular Ca2+ concentration. The response to the agonists was marginal in the absence of glycine, and diminished by Mg2+ and NMDA receptor antagonists. Furthermore, exposure to agonists of ZE1-1 cells expressing the epsilon1/zeta1 NMDA receptor channel resulted in the release of lactate dehydrogenase (LDH) activity in the culture medium indicating agonist-induced cell death. NMDA receptor antagonists inhibited the LDH activity release. These results suggest that ZE1-1 cells will provide a useful screening system for novel drugs acting on the epsilon1/zeta1 NMDA receptor channel.

MTT colorimetric assay system for the screening of anti-orthomyxo- and anti-paramyxoviral agents.

A rapid and sensitive method was developed for screening potential antiviral agents against orthomyxo- and paramyxoviruses, using the MTT method with cell culture suspensions. The cell lines used for the assay were as follows: MDCK cells for the influenza A virus (Fluv. A), HeLa cells for the respiratory syncytial virus (RSV), and Vero cells for the measles virus (MSV). Test compounds were diluted and plated in 96-well round-bottomed microtiter plates. Trypsinized cell suspensions and viruses were added to each well, the plates were then centrifuged (700 x g, 5 min, room temperature), and incubated for several days. The MTT assay was carried out after the degeneration of virus-infected cells became evident. The optical density (OD) of formazan was determined using a computer-controlled microplate reader. With this assay system, the EC50 values of Ribavirin (used as the reference compound) were 3.7 micrograms/ml for Fluv. A, 4.5 micrograms/ml for RSV, and 12.3 micrograms/ml for MSV, respectively. These EC50 values were equivalent to those obtained using the plaque reduction assay. The confluent cell culture system was inadequate for antiviral assays against RSV and MSV when the MTT method was used, because the inhibition of formazan formation was not observed in viral-infected cells. Moreover, the suspension method is more sensitive to the cytotoxicity of antiviral agents than the confluent cell culture system.

Use of lactate dehydrogenase to evaluate the anti-viral activity against influenza A virus.

The detection of lactate dehydrogenase (LDH) can be used to evaluate efficiently anti-influenza A virus agents. LDH levels in the virus-infected Madin-Darby canine kidney cell cultures were significantly higher than in controls, were in proportion to the degree of virus infection, and corresponded to a decrease in mitochondrial dehydrogenase activity as assayed using a tetrazolium colorimetric assay (MTT method). The EC50 value and cytotoxicity of ribavirin, 3-deazaguanine, pyrazofurin, and carbodine against influenza A virus as measured by the LDH detection method was equivalent to that derived by the MTT method.

Mouse model of respiratory syncytial virus infection to evaluate antiviral activity in vivo.

In this study, we have developed a practical mouse model for evaluating in vivo the antiviral activity of compounds against respiratory syncytial virus (RSV) infection. BALB/c mice are not particularly susceptible to RSV infection; however, infection rates were improved by pretreatment with the immunosuppressive agent cyclophosphamide (CYP). When mice were inoculated intranasally with RSV A2 strain, the pulmonary RSV titres of CYP-pretreated 10-week-old mice were higher than those of untreated 10-week-old and 28-week-old mice, peaking on days 4 and 5 post-infection. Sections of lung from RSV-infected mice pretreated with CYP, taken on day 4 post-inoculation, showed widespread evidence of interstitial pneumonia and other significant pathological changes. We also confirmed that ribavirin, a representative antiviral agent, significantly reduced the pulmonary RSV titres of mice pretreated with CYP when administered intraperitoneally.

Range of motion after Bankart repair. Vertical compared with horizontal capsulotomy.

We studied the range of shoulder motion of patients who underwent vertical as compared with horizontal capsulotomies during open Bankart repair for recurrent anterior dislocations of the shoulder. A vertical capsulotomy was used in 10 shoulders and a horizontal capsulotomy was used in 14 shoulders. Except for the method of capsulotomy, the surgical procedure and postoperative rehabilitation were the same. The range of motion was measured at 1.5, 2, 3, 4, 5, 6, 9, and 12 months after the surgery, and at the final follow-up (average, 49 months for the vertical and 26 months for the horizontal group). No dislocations recurred, and the anterior apprehension test was negative in all of the patients in both groups. External rotation in abduction was greater in the horizontal group than in the vertical group; the differences were significantly greater at 9 months and 12 months after surgery and at the final follow-up. External rotation in adduction, flexion, and internal rotation were not significantly different between the groups. We conclude that Bankart repair through a horizontal capsulotomy preserves a better range of external rotation in abduction than does a vertical approach.

A rare anatomic variant of the superior glenohumeral ligament.

The attachment of the superior glenohumeral ligament (SGHL) to the upper pole of the glenoid is variable and 3 types have been described. We report an anatomic variant of SGHL attachment to the upper pole of the glenoid that has not heretofore been reported in the literature. In this case, the SGHL overrode the biceps origin, continued to the superior labrum posteriorly, and had no attachment to the middle glenohumeral ligament or the anterior labrum. This variant was detected during routine arthroscopic examination undertaken before surgery on a rotator cuff tear.

Influence of the varus-valgus instability on the contact of the femoro-tibial joint.

The varus-valgus instability of the knee joint is mainly due to ruptured or lax collateral ligaments. The purpose of this investigation was to study the influence of the varus-valgus instability on the contact pressures of the femoro-tibial joint. Six fresh knee specimens of human cadavers were tested to measure the contact pressure on the tibia plateau of the knee joint at varus or valgus alignment under various loads and at full extension. Pressure transducers and Bourdon tube pressure gauges were used simultaneously for recording pressure. At neutral alignment of the knee with the menisci intact, the peak pressure increased linearly with forces up to 4 MPa. With increasing varus alignment, the peak contact pressure on the medial plateau not covered by the menisci increased up to a maximum of 7.3 MPa at 5 degrees varus, and at 5 degrees valgus, the peak pressure on the lateral plateau was 7.8 MPa. After total meniscectomy, the contact pressure increased up to a maximum of 7.4 MPa at a force of 2700 N. With increasing varus alignment, the contact pressure on the medial plateau increased to 8.1 MPa at 5 degrees varus and on the lateral plateau 9.2 MPa at 5 degrees valgus.

[A case with severe neurological involvement due to vitamin B1 deficiency associated with megaduodenum].

We reported a case of 4-year-old boy with multiple vitamin deficiencies, especially vitamin B1 deficiency. He had megaduodenum associated with membranous stenosis on upper jejunum. He showed recurrent vomiting at his infantile period, and recently intermittent neurological symptoms. When he was admitted to our hospital, he could not walk and showed masked face, absent deep tendon reflexes, horizontal and vertical nystagmus, proximally dominant muscle weakness and multiple vitamin deficiencies. Oral administration of small doses of vitamin B1 (20 mg/day) could make remarkable clinical improvements. At three weeks after the treatment he could walk and run. Before the admission he had febrile convulsions and showed transiently striatal low density on CT image. We concluded that his neurological symptoms were due to vitamin B1 deficiency associated with megaduodenum. When a patient with intestinal anomaly shows neurological symptoms, we should think of vitamin deficiency.

[Study on identification of a calcium pyrophosphate dihydrate crystal using electron diffraction technique].

Electron diffraction technique was used to identify a single crystal in synovial fluid and tissue obtained from patients with pseudogout. One hundred and ten kinds of reciprocal lattice planes of the triclinic calcium pyrophosphate dihydrate (CPPD) crystal were made from its crystallographical data. Electron diffraction patterns obtained from synthesized crystal closely fitted to the reciprocal lattice planes. Synovial fluid of ten patients who had chondrocalcinosis in the knee joint were examined. Diffraction patterns of a single rhomboid-shaped crystal fitted to the reciprocal lattice planes and these kinds of crystals were identified as triclinic CPPD crystals. A single triclinic crystal in the calcified meniscus prepared for transmission electron microscopy was identified in the same way as above. On the other hand, there weren't enough crystallographical data on the monoclinic CPPD crystal to construct the reciprocal lattice planes. Instead of comparing diffraction patterns with the reciprocal lattice planes, diffraction patterns of a single rod-like crystal were compared with those of a synthesized monoclinic CPPD crystal. This comparison combined with energy dispersive elemental analysis made it possible to identify a single rod-like crystal as a monoclinic CPPD crystal.

[Histological study on radiosensitivity of cervical carcinoma with special reference to 3 subtypes of squamous cell carcinoma].

Morphological investigation were made on the radiosensitivity, according to the histological types; keratinizing(K), large cell nonkeratinizing(LNK), and small cell nonkeratinizing(S) type. The results obtained are as follows. Histological and cytological estimation of irradiated effects were examined at the 7th day after 10 Gray test-irradiation: (i) Histological studies were done with the biopsy specimens (76 cases): There were significant differences among 3 histological types (LNK greater than K greater than S, P less than 0.05). (ii) Cytological studies were done as well as histological ones (48 cases): There was a significance between two groups (LNK greater than S, K greater than S, P less than 0.05). (iii) There was a correlation between the histological and cytological irradiation effects. DNA histogram patterns were also examined spectrophotometrically. It seemed that the patterns after test-irradiation was broad and these peaks were situated rather to high ploidy in histologically and cytologically highly effective cases. Early ultrastructural findings after test-irradiation were as follows: (i) appearance of intra-nuclear "pseudoinclusion", (ii) swelling of mitochondria, endoplasmic reticulum, and so on. Although these characteristics presented the same sequence of events without regard to their histological types, they were not synchronized. It seemed that they appeared earlier after test-irradiation in histologically high-sensitivity cases than in low-sensitivity cases.

A colorimetric LDH assay for the titration of infectivity and the evaluation of anti-viral activity against ortho- and paramyxoviruses.

A rapid and precise screening assay was developed for in vitro evaluation of anti-orthomyxo- and anti-paramyxovirus agents. The procedure is spectrophotometrical assessment for viability of cells via extracellular leakage of lactic dehydrogenase (LDH). HMV-II cells, a human melanoma cell line was found to be suitable for the titration of virus infectivity and screening of anti-viral agents for orthomyxo- and paramyxoviruses. Comparative titration of infectivity of stock viruses by the LDH and the MTT in site reduction of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) methods with HMV-II cells as well as plaque titration with MDCK, Vero and HeLa cells was carried out. The LDH method was comparable or more sensitive for influenza viruses (FLUV)-A, B, C, parainfluenza viruses (PFLUV)-1, 2 and less sensitive for PFLUV-3, mumps virus (MPSV), measles viruses (MLSV) and respiratory syncytial virus (RSV) than the plaque titration. The 50% effective concentration (EC50) of 1-beta-D-ribofuranosyl-1, 2, 4-triazol-3-carboxamide (ribavirin) and 5-ethynyl-1-beta-D-ribofuranosyl-imidazole-4-carboxamide (EICAR) against orthomyxo- and paramyxoviruses were examined comparatively by the LDH, MTT and plaque reduction (PR) methods. The EC50 values of FLUV-C and PFLUV-1 were able to be evaluated only by the LDH but not by the MTT and PR methods. The LDH method with HMV-II cells simplifies the assay procedure and permits the evaluation of a large number of compounds for anti-orthomyxo- and anti-paramyxoviruses activity in vitro.