Comparable evaluation of serological diagnostic tests (ELISA, IFA and PA methods) for the detection of anti-Borrelia burgdorferi antibody]

We have evaluated the usefulness of the enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibody assay (IFA) and particle agglutination (PA) method as serological screening tests for Lyme-borreliosis. Serum samples obtained from two patients with Lyme-borreliosis showed marked high antibody titers for Borrelia burgdorferi when measured by these methods. Of the serum of 368 healthy members of the Self-Defense Force in north-eastern Japan screened for the antibody to B. burgdorferi, 8.4%, 3.7%, 4.6% were found positive by the ELISA, IFA, and PA method, respectively. However, Western blot analysis of these "positive" sera demonstrated no identical bands to those seen in the serum from the patients with Lyme-borreliosis. While 85% and 15% of Treponema pallidum hemagglutination test (TPHA)-positive sera (20 samples) showed a false-positive reaction by the ELISA and IFA method, respectively, no cross-reaction to the anti-B. burgdorferi antibody was observed in these sera by the PA method. The analysis of the serum of the patients with autoimmune diseases (rheumatoid arthritis; 11 cases, systemic lupus erythematosus; 46 cases) by the ELISA and PA methods resulted in a cross-reaction to some extent, which suggested that the antibodies produced by autoimmune mechanisms such as the anticardiolipin antibody can cause a cross-reaction to the anti-B. burgdorferi antibody. These findings indicate that the PA and ELISA rather than the IFA method should be recommended for rapid and conventional screening of Lyme-borreliosis and that serum "positive" for the anti-B. burgdorferi antibody determined by these tests should be confirmed by Western blot analysis to negate the cross-reactions.     

Assimilatory reduction of nitrate in Rhizobium meliloti

Nitrate and nitrite reductases in the crude extract of aerobically grown Rhizobium meliloti were determined with methylviologen as electron donor at pH 7. Nitrate reductase was detected in the cells grown in the medium that did not contain nitrate, and in the presence of nitrate the specific activity increased about 2-fold. Nitrite reductase was induced by nitrate and produced ammonia from nitrite. In nitrate reducing cells, two kinds of O₂ labile nitrate reductase were found. One enzyme had optimal pH at 7 and was stabilized to O₂ by treating with DEAE-Toyopearl 650M. The other had optimal pH at 9 and was stabilized by the addition of dithiothreitol and EDTA. Nitrate reductase stabilized by DEAE-Toyopearl 650M treatment was purified 3,360-fold from crude extract. The purified enzyme showed a single protein band in polyacrylamide gel electrophoresis, and there was no absorption peak in the visible region. It had a molecular weight of 64,000 in SDS PAGE and 58,000 on Sephadex G-100 gel filtration. K_m for nitrate was 0.9 mM. It was inhibited by p-chloromercuribenzoate, cyanide, and α,α'-dipyridyl.     

Early post‐operative intravenous tacrolimus in pediatric liver transplant recipients is not superior to oral tacrolimus

We aimed to compare the early results of i.v. with p.o. TAC as a primary immunosuppressant in pediatric patients undergoing LT. This retrospective study enrolled 75 children who underwent LT and received TAC‐steroid regimens as a primary immunosuppressant between September 2011 and October 2015 at our institution. Thirty‐five recipients received TAC i.v. and 40 received TAC p.o. Early results were evaluated and compared, including ACR, EBV, or CMV infection; renal adverse effects; and hospital stay. Comparisons of 90‐day post‐transplant results showed that the rates of overall viral (74% vs 40% P < 0.002), EBV (46% vs 17.5% P < 0.008), and CMV (51% vs 30% P = 0.05) infections were significantly higher in the i.v. than in the p.o. group. Neither regimen has any adverse effects on renal function. There were no between‐group differences in ACR incidence and severity, serum creatinine concentration, and hospital stay. Patient and graft survival rates at 3 months and 1 year did not differ significantly between the two groups. Compared with p.o. treatment, i.v. administration of high TAC concentration did not have beneficial post‐transplant effects on ACR incidence and severity, while increasing the incidence of viral infections in pediatric LT.     

Management of patients with coexisting obstructive sleep apnea and laryngopharyngeal reflux disease

Obstructive sleep apnea syndrome (OSAS) and Laryngopharyngeal reflux disease (LPR) are both common health problems causing severe morbidity. Since they have similar risk factors, the prevalence of LPR among patients with OSAS is higher compared with general population. However, there exist only a few studies showing the potential causal relation between LPR and OSAS. The aim of this study was to evaluate the coexistence between OSAS and LPR and to determine whether the therapy of OSAS alters LPR parameters and vice versa. In this study, 44 patients underwent double probed 24?h pH monitoring simultaneously with polysomnography due to the complaints of obstructive sleep apnea and reflux. Twenty of those 44 patients were diagnosed with both OSAS and LPR. Among those patients, 10 patients with mild to moderate OSAS were given only LPR treatment for 3?months. The remaining 10 patients who had severe OSAS underwent CPAP treatment for 3?months. After the end of treatment, all patients were reevaluated with double probed 24?h pH monitoring simultaneously with PSG. Moreover, the patients were evaluated subjectively by Epworth Sleepiness Scale (ESS), snoring Visual Analogue Scale (VAS), Reflux Symptom Index (RSI), and Reflux Finding Score (RFS). The results of this study revealed that OSAS and LPR coexist frequently. LPR treatment did not improve the polysomnographic parameters, but significantly reduced ESS and snoring VAS (p?=?0.02 and p?=?0.007, respectively). Although the CPAP treatment significantly improved subjective parameters of reflux, such as RSI and RFS (p?=?0.016 for both), there was no significant improvement in objective parameters of 24-h pH monitoring. We concluded that since there is a high frequency of coexistence between LPR and OSAS, all patients with OSAS should also be queried for LPR symptoms. In addition, more in-depth and comprehensive research is required to elucidate the association between OSAS and LPR.     

PRODUCTION OF ALPHA-FETOPROTEIN BY HUMAN GERM CELL TUMORS IN VIVO AND IN VITRO

The production of alpha-fetoprotein (AFP) by human germ cell tumors was studied in surgical specimens, cultured cells and transplanted tumors in nude mice. AFP was detected most frequently in yolk sac (endodermal sinus) tumors, and it was also detected, though only occasionally, in embryonal carcinomas and in teratomas. AFP-positive cells in teratomas presented a glandular or hepatoid appearanece. Our experimental data suggest that AFP production in embryonal carcinomas is due to the functional expression of yolk sac differentiation preceding its morphologic expression. Although the transplanted tumors maintained the productivity of AFP, the cultured cells were not successful in maintaing a continuous AFP production.     

Effect of solvent evaporation rate on microphase-separated structure of segmented poly(urethane-urea) prepared by solution casting

A useful instrument for polymer film preparation by solution casting was employed in this study. It enabled us to control the solvent evaporation rate of the polymer solution. By using this instrument, the aggregation of hard segments in segmented poly(urethane-urea) (SPUU) was investigated. SPUU was prepared from poly(tetramethy1ene oxide), 4,4′-diphenylmethane diisocyanate and ethylenediamine. The effect of solvent evaporation rate on the microphase-separated structure of SPUU was elucidated by dynamic mechanical analysis, tensile test, differential scanning calorimetry analysis, small-angle X-ray scattering measurement, IR and IR dichroism analyses. The aggregation of hard segments in SPUU was observed to be affected considerably by the solvent evaporation rate of the cast film during the preparation. It was found that the slower the solvent evaporation rate, the higher the aggregation of hard segments to form rigid hard segment domains in SPUU. Nine months after casting, this casting effect still remained on the aggregation state of hard segments of SPUU films, although the interdomain spacing was not influenced by its rate.     

Extracellular matrix dynamics in cell migration,invasion and tissue morphogenesis

This review describes how direct visualization of the dynamic interactions of cells with different extracellular matrix microenvironments can provide novel insights into complex biological processes. Recent studies have moved characterization of cell migration and invasion from classical 2D culture systems into 1D and 3D model systems, revealing multiple differences in mechanisms of cell adhesion, migration and signalling—even though cells in 3D can still display prominent focal adhesions. Myosin II restrains cell migration speed in 2D culture but is often essential for effective 3D migration. 3D cell migration modes can switch between lamellipodial, lobopodial and/or amoeboid depending on the local matrix environment. For example, “nuclear piston” migration can be switched off by local proteolysis, and proteolytic invadopodia can be induced by a high density of fibrillar matrix. Particularly, complex remodelling of both extracellular matrix and tissues occurs during morphogenesis. Extracellular matrix supports self‐assembly of embryonic tissues, but it must also be locally actively remodelled. For example, surprisingly focal remodelling of the basement membrane occurs during branching morphogenesis—numerous tiny perforations generated by proteolysis and actomyosin contractility produce a microscopically porous, flexible basement membrane meshwork for tissue expansion. Cells extend highly active blebs or protrusions towards the surrounding mesenchyme through these perforations. Concurrently, the entire basement membrane undergoes translocation in a direction opposite to bud expansion. Underlying this slowly moving 2D basement membrane translocation are highly dynamic individual cell movements. We conclude this review by describing a variety of exciting research opportunities for discovering novel insights into cell‐matrix interactions.     

A rare case of pulmonary typical carcinoid with prominent acinic cell differentiation,resembling acinic cell carcinoma

We herein describe a rare case of low‐grade endobronchial tumor that exhibited two distinct features of typical carcinoid and acinic cell carcinoma (ACC) by immunohistochemical and ultrastructure study. ACC was suspected on transbronchial biopsy. The resected specimen showed that the tumor surface comprised an acinic cell component (40% of the tumor), and the central area comprised typical carcinoid (60% of the tumor). The acinic cell component was positive for chromogranin A, synaptophysin and alpha‐1‐antichymotrypsin. Additionally, this component showed focal apical membranous staining for DOG1 and weak positivity for BCL10 and SOX10. Conversely, the carcinoid component was negative for all proteins except for chromogranin A and synaptophysin. Electron microscopy indicated zymogen‐type granules (600–800 nm in diameter) in the acinic cell component, whereas neuroendocrine‐type granules (200–300 nm in diameter) were observed in the carcinoid component. Nuclear NR4A3 immunostaining, which is highly specific for ACC of the salivary gland, was negative in this case. We conclude that the pulmonary carcinoid tumor with true zymogen‐type granules could be seen but showed superficial similarities to ACC based on negative nuclear staining for NR4A3. Pulmonary carcinoids encompass a wide morphological spectrum and may exhibit prominent acinic cell differentiation.