Search

In this work, we considered the autoantibodies proposed as putative biomarkers of demyelination taking into account their reactivity towards myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP). These myelin proteins are among the most commonly researched targets in the immunopathology of demyelinating diseases. In this context, the development of assays for autoantibody detection can contribute as a predictive value for the early diagnosis of demyelinating diseases. Hence, we aimed to address the application of silver nanoparticles (AgNPs) as a sensing device of autoantibodies. AgNPs were synthesized via a chemical reduction method and characterized using atomic force microscopy (AFM), X-ray diffractometry, dynamic light scattering, and UV-visible spectrophotometry. The process of peptide conjugation on the nanoparticles was also analyzed. The autoantibody recognition by the peptide-conjugated AgNPs was evaluated with UV-visible spectrophotometry, atomic force spectroscopy (AFS), and color changing. AgNPs exhibited spherical morphology, low polydispersity, face-centered cubic crystal structure, and an average size of 29.3±3.0 nm. The hydrodynamic diameter variation and AFM showed that the MBP peptide induced greater agglomeration, compared to MOG peptide. The AFS measurements indicated the efficient binding of peptides to the AgNPs maintaining their activity, revealed by typical adhesion force and shapes of curves. The absorption spectrum features were more affected by the interaction with the specific autoantibodies, which also caused a visible color change in suspension providing a qualitative response. We described a preliminary study of MOG- and MBP-conjugated AgNPs which showed to be applicable in the autoantibody recognition. These have promising implication in the searching for biological markers for diagnostic purposes in the demyelination context, in which the nanoscale sensing exploitation is recent.

Made available in DSpace on 2020-12-10T19:54:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) National Institute for Science and Technology on Organic Electronics(INEO) Multiple Sclerosis (MS) is a chronic inflammatory disorder in the central nervous system for which biomarkers for diagnosis still remain unknown. One potential biomarker is the myelin basic protein. Here, a nanoimmunosensor based on atomic force spectroscopy (AFS) successfully detected autoantibodies against the MBP85-99 peptide from myelin basic protein. The nanoimmunosensor consisted of an atomic force microscope tip functionalization with MBP85-99 peptide, which was made to interact with a mica surface coated either with a layer of anti-MBP85-99 (positive control) or samples of cerebrospinal fluid (CSF) from five multiple sclerosis (MS) patients at different stages of the disease and five non-MS subjects. The adhesion forces obtained from AFS pointed to a high concentration of anti-MBP85-99 for the two patients at early stages of relapsing-remitting multiple sclerosis (RRMS), which were indistinguishable from the positive control. In contrast, considerably lower adhesion forces were measured for all the other eight subjects, including three MS patients with longer history of the disease and under treatment, without episodes of acute MS activity. We have also shown that the average adhesion force between MBP85-99 and anti-MBP85-99 is compatible with the value estimated using steered molecular dynamics. Though further tests will be required with a larger cohort of patients, the present results indicate that the nanoimmunosensor may be a simple tool to detect early-stage MS patients and be useful to understand the molecular mechanisms behind MS. Univ Fed Sao Carlos, Dept Phys Chem & Math, Nanoneurobiophys Res Grp, BR-18052780 Sorocaba, SP, Brazil Univ Sao Paulo, Inst Trop Med, BR-05403000 Sao Paulo, SP, Brazil Sao Paulo State Univ, Dept Neurol Psychol & Psychiat, BR-18618687 Botucatu, SP, Brazil Univ Sao Paulo, Sao Carlos Inst Phys, BR-13560970 Sao Carlos, SP, Brazil Univ Sao Paulo, Inst Chem, Dept Biochem, BR-05508000 Sao Paulo, SP, Brazil Rheabiotech Lab Res & Dev, BR-13084791 Campinas, SP, Brazil Pontifical Catholic Univ Sao Paulo, BR-18030010 Sorocaba, SP, Brazil Sao Paulo State Univ, Dept Neurol Psychol & Psychiat, BR-18618687 Botucatu, SP, Brazil FAPESP: 2013/14262-7 FAPESP: 2014/21530-0 FAPESP: 2015/14360-4 FAPESP: 2015/06847-0 FAPESP: 2015/36143-2 FAPESP: 2015/052836 FAPESP: 2014/12082-4 FAPESP: 2012/50839-4 FAPESP: 2014/50869-6 CNPq: 465572/2014-6 CNPq: 308658/2015-9 CNPq: 459768/2014-0 CAPES: 23038.000776/201754

Conjugates based on cell-penetrating peptides (CPPs) are scientifically relevant owing to their structural complexity; their ability to enter cells and deliver drugs, labels, antioxidants, bioactive compounds, or DNA fragments; and, consequently, their potential for application in research and biomedicine. In this study, carboxyamidated fluorescently labeled conjugates FAM-GG-TAT(47-57)-NH2 and FAM-PEG6-TAT(47-57)-NH2 and photosensitizer-labeled conjugate Chk-PEG6-TAT(47-57)-NH2 [where TAT(47-57) is the CPP, 5(6)-carboxyfluorescein is the (FAM) fluorophore, chlorin k (Chk) is the photosensitizer, and the dipeptide glycyl–glycine (GG) or hexaethylene glycol (PEG6) is the spacer] were originally designed, prepared, and fully characterized. Practically, all chemical reactions of the synthetic steps (peptide synthesis, spacer incorporation, and conjugation) were microwave-assisted at 60 °C using optimized protocols to give satisfying yields and high-quality products. Detailed analyses of the conjugates using spectrofluorimetry and singlet oxygen detection showed that they display photophysical properties typical of FAM or Chk. Anticandidal activity assays showed that not only this basic property of TAT(47-57) was preserved in the conjugates but also that the minimal inhibitory concentration was slightly reduced for cells incubated with PS-bearing conjugate Chk-PEG6-TAT(47-57)-NH2. Overall, these results indicated that the synthetic approach on-resin assisted by microwaves at 60 °C is simple, straightforward, selective, metal-free, sufficiently fast, cleaner, and more cost-effective than those previously used for preparing this type of macromolecule. Furthermore, such new data show that microwaves at 60 °C and/or conjugation did not harm the integrity of the conjugates’ constituents. Therefore, FAM-GG-TAT(47-57)-NH2, FAM-PEG6-TAT(47-57)-NH2, and Chk-PEG6-TAT(47-57)-NH2 have high potential for practical applications in biochemistry, biophysics, and therapeutics.

This paper describes the protection system of MON.ITA HVDC Link in all its parts, which has been designed taking into account the different configuration possibilities. AC protection relays, including AC connections, filters and transformers 400/205/205 kV and DC protection relays are presented with a functional description of the entire protection system. Regarding DC part, the protections of AC/DC conversion system, DC Cables and Common Neutral system are reviewed. General protection criteria, adopted schemes to manage main critical events and operating sequences to stop AC/DC conversion, depending on the severity of the fault, are explained.

Purpose Cardiopulmonary bypass (CPB) and extracorporeal membrane oxygenation (ECMO) may be used as extracorporeal circulation (ECC) during lung transplantation. We aimed to compare CPB and ECMO in terms of short-term outcomes in lung transplantation (LTx). Methods Among 185 patients undergoing LTx including 81 living-donor-related LTx from 2008 to June, 2018 in our institution, CPB was used in 41 patients and ECMO in 89 patients. Since 2013, we have routinely used ECMO, while CPB has been selectively used in pediatric and complicated cases requiring PA replacement or ASD closure. We retrospectively evaluated perioperative factors and short-term outcomes including delayed chest closure (DCC), in comparison of CPB group and ECMO group. DCC is required due to primary graft dysfunction manifesting as hypoxia, pulmonary hypertension, and lung edema. Assuming that CPB causes systemic inflammation leading to systemic edema, we also investigated perioperative increase ratio of body weight. Results Gender, surgical procedure (single or bilateral), and donor (cadaveric or living) and indications were not significantly different between the two groups, while patients in ECMO group were significantly older (40 vs 32 years, p=0.038), and weighed more (47 vs 38 kg, p=0.001). Thirty-day mortality was similar (1.1% in ECMO group; 2.4% in CPB group). The operation time, ECC time, and the amount of blood loss were not significantly different, while ECMO group was associated with significantly less transfusion intraoperatively (4339 ml in ECMO group; 6753 ml in CPB group, p=0.002). Reoperations for bleeding were less frequently required in ECMO group than in CPB group (7% vs 29%, p=0.003). DCC was less frequently required in ECMO group than in CPB group (9% vs 29%, p=0.013). The increase of body weight measured at the time of ICU admission was significantly remarkable in CPB group than in ECMO group (increased by 9.7% vs by 4.7%, p=0.006). Conclusion LTx using ECMO was associated with less intraoperative transfusion, and less postoperative re-exploration to CPB. ECMO was associated with less frequent delayed chest closure, possibly related to less systemic edema.

Aim A Kampo medicine, kososan (KS), has been used clinically for the treatment of certain depression-like symptoms. Our previous studies using the stress-induced depression-like model mice, showed that oral KS treatment leads to an antidepressive-like effect via the normalization of dysfunction of hypothalamic-pituitary-adrenal axis. In the present study, proteomic analysis was used to identify brain hypothalamus proteins that are affected by KS treatment. Methods KS was orally administered to stress-induced depression-like model mice at 1.0 g/kg/day for 9 days, and the hypothalamus was then analyzed using agarose 2-D gel electrophoresis, followed by mass spectrometry-based protein identification, and then western blot analysis and immunohistochemical analysis. Results The expression of eight proteins was increased or decreased in the hypothalamus of the model mice, but recovered on oral KS treatment. Among them, metabotropic glutamate receptor 2 (mGluR2) and 2′,3′-cyclic nucleotide 3′-phosphodiesterase 1 (CNPase1) were identified as depression-related proteins. Western blot of the hypothalamus confirmed these results and showed that the expression of CNPase2 was opposite to that of CNPase1. On immunohistochemistry, mGluR2 expression in the hypothalamus paraventricular nucleus was downregulated and CNPase2 expression was upregulated in the cerebral fornix in the model mouse, but KS returned them to normal. Conclusion mGluR2 and CNPase in the hypothalamus are associated with the antidepressive-like activity of KS.

SUMMARY Background and objectives The Kidd blood group system consists of polymorphic antigens, Jka (JK1) and Jkb (JK2), and a high-incidence antigen, Jk3. Anti-Jk3 is often observed in immunised Jk(a−b−) individuals. In this study, we aimed to establish a human hybridoma cell line secreting monoclonal anti-Jk3 (HIRO-294). Materials and methods Peripheral blood lymphocytes of a Filipino woman with the Jk(a−b−) phenotype having anti-Jk3 were transformed with Epstein-Barr virus and then hybridised with the myeloma cell line JMS-3 using the polyethylene glycol (PEG) method. The reactivity and specificity of the anti-Jk3 were examined by serology and flow cytometry. Results Four hybridoma clones secreting anti-Jk3 were established and the antibody from one of these clones, HIRO-294, was examined. The reactivity of HIRO-294 was positive with 227 Jk(a+b−) red blood cells (RBCs), 298 Jk(a−b+) RBCs, and 1043 Jk(a+b+) RBCs, but was negative with 21 Jk(a−b−) RBCs. Eluates from Jk(a+b−) RBCs and Jk(a−b+) RBCs sensitised with the anti-Jk3 were cross-reacted with Jk(a−b+) RBCs and Jk(a+b−) RBCs, respectively. The reactivity of HIRO-294 was enhanced by the treatment of RBCs with ficin, trypsin, pronase and α-chymotrypsin, but was not changed by their treatment with neuraminidase, dithiothreitol and ethylenediaminetetraacetic acid (EDTA) glycine acid (GA). The RBCs sensitised by the anti-Jk3 were not agglutinated with the commercial reagents of anti-Jka and anti-Jkb by saline test, whereas the nonsensitised RBCs or those sensitised by monoclonal anti-D [HIRO-3, immunoglobulin G (IgG) class] were agglutinated with those reagents. Conclusions We established a human hybridoma cell line secreting monoclonal anti-Jk3 (HIRO-294). This antibody had unique specificity, recognising the Kidd glycoprotein including the Jka/Jkb polymorphic site.

Objective:Insufficient sleep or short sleep duration is a risk factor for hypertension associated with cardiovascular disease in adults. Irregular sleep-wake rhythm is common problems among university students. We investigated the effect of sleep-wake rhythm or sleep duration on endothelial function

Purpose The perioperative survival of lung transplantation (LTx) for patients with idiopathic pulmonary arterial hypertension is known to be worse than for those with other underlying diseases because of difficulties in managing perioperative haemodynamic instability. High dose epoprostenol (PGI2) is frequently used for patients with end-stage pulmonary hypertension (PH) before transplantation, however, it is generally discontinued at the time of transplantation. We hypothesized that tapering epoprostenol rather than abrupt discontinuation after reperfusion would improve perioperative outcomes. Methods We performed 193 LTxs since 2008 and 16 PH patients were on high dose epoprostenol (10-200 ng/kg/min) therapy. Epoprostenol was discontinued at the time of lung transplantation in 6 patients and it was tapered after reperfusion in ten patients. In the Discontinued group, epoprostenol was discontinued intraoperatively during extracorporeal membrane oxygenation (ECMO) or cardiopulmonary bypass and was not resumed postoperatively. In the Tapered group, epoprostenol was discontinued during extracorporeal support and was resumed at the quarter of preoperative dose after reperfusion. It was then gradually tapered typically over the following two weeks according to the heart function on echocardiography. Results One patient in the Discontinued group died on day 125 due to multiple organ failure, in contrast, no operative death was encountered in the Tapered group. Delayed chest closure and postoperative ECMO were more frequently required in the Discontinued group than the Tapered group (p = 0.0076, respectively). Primary graft dysfunction score was significantly lower in the Tapered group than the Discontinued group (0.6 ± 1.3 vs 2.2 ± 1.3 at 0h, p = 0.031; 0.4 ± 0.8 vs 2.0 ± 1.5 at 24h, p = 0.028; 0.3 ± 0.7 and 1.8 ± 1.5 at 48h, p = 0.033). Furthermore, the Tapered group required significantly shorter duration of postoperative mechanical ventilation (15.1 ± 6.8 vs 41.5 ± 41.9 days, p = 0.039). Conclusion Tapering epoprostenol administration after reperfusion for PH may be a valuable new strategy which is associated with better perioperative outcomes.

Neural circuits that undergo reorganization by newborn interneurons in the olfactory bulb (OB) are necessary for odor detection and discrimination, olfactory memory, and innate olfactory responses, including predator avoidance and sexual behaviors. The OB possesses many interneurons, including various types of granule cells (GCs); however, the contribution that each type of interneuron makes to olfactory behavioral control remains unknown. Here, we investigated the in vivo functional role of oncofetal trophoblast glycoprotein 5T4, a regulator for dendritic arborization of 5T4 -expressing GCs (5T4 GCs), the level of which is reduced in the OB of 5T4 knock-out (KO) mice. Electrophysiological recordings with acute OB slices indicated that external tufted cells (ETCs) can be divided into two types, bursting and nonbursting. Optogenetic stimulation of 5T4 GCs revealed their connection to both bursting and nonbursting ETCs, as well as to mitral cells (MCs). Interestingly, nonbursting ETCs received fewer inhibitory inputs from GCs in 5T4 KO mice than from those in wild-type (WT) mice, whereas bursting ETCs and MCs received similar inputs in both mice. Furthermore, 5T4 GCs received significantly fewer excitatory inputs in 5T4 KO mice. Remarkably, in olfactory behavior tests, 5T4 KO mice had higher odor detection thresholds than the WT, as well as defects in odor discrimination learning. Therefore, the loss of 5T4 attenuates inhibitory inputs from 5T4 GCs to nonbursting ETCs and excitatory inputs to 5T4 GCs, contributing to disturbances in olfactory behavior. Our novel findings suggest that, among the various types of OB interneurons, the 5T4 GC subtype is required for odor detection and discrimination behaviors. SIGNIFICANCE STATEMENT Neuronal circuits in the brain include glutamatergic principal neurons and GABAergic interneurons. Although the latter is a minority cell type, they are vital for normal brain function because they regulate the activity of principal neurons. If interneuron function is impaired, brain function may be damaged, leading to behavior disorder. The olfactory bulb (OB) possesses various types of interneurons, including granule cells (GCs); however, the contribution that each type of interneuron makes to the control of olfactory behavior remains unknown. Here, we analyzed electrophysiologically and behaviorally the function of oncofetal trophoblast glycoprotein 5T4, a regulator for dendritic branching in OB GCs. We found that, among the various types of OB interneuron, the 5T4 GC subtype is required for odor detection and odor discrimination behaviors.

The objective of this study was to compare the sensitivity of detection of lung nodules on low-dose screening CT images between radiologists and technologists.11 radiologists and 10 technologists read the low-dose screening CT images of 78 subjects. On images with a slice thickness of 5 mm, there were 60 lung nodules that were ≥5 mm in diameter: 26 nodules with pure ground-glass opacity (GGO), 7 nodules with mixed ground-glass opacity (GGO with a solid component) and 27 solid nodules. On images with a slice thickness of 2 mm, 69 lung nodules were ≥5 mm in diameter: 35 pure GGOs, 7 mixed GGOs and 27 solid nodules. The 21 observers read screening CT images of 5-mm slice thickness at first; then, 6 months later, they read screening CT images of 2-mm slice thickness from the 78 subjects.The differences in the mean sensitivities of detection of the pure GGOs, mixed GGOs and solid nodules between radiologists and technologists were not statistically significant, except for the case of solid nodules; the p-values of the differences for pure GGOs, mixed GGOs and solid nodules on the CT images with 5-mm slice thickness were 0.095, 0.461 and 0.005, respectively, and the corresponding p-values on CT images of 2-mm slice thickness were 0.971, 0.722 and 0.0037, respectively.Well-trained technologists may contribute to the detection of pure and mixed GGOs ≥5 mm in diameter on low-dose screening CT images.

Alpha-hydroxy acids (AHAs), primarily glycolic and lactic acids, are widely used in cosmetics to alleviate dyspigmentation, photodamage, and other aging skin conditions and as pH adjusters. Glycolic acid reportedly enhances skin damage after repeated ultraviolet light exposure, e.g., increased sunburn cell formation. This study assessed potential in vitro skin penetration of lactic acid and malic acid incorporated into rinse-off personal care products, compared with rinse-off and leave-on exposures to glycolic acid (10%, pH 3.5) in a reference lotion. Radiolabeled AHA-fortified shampoo, conditioner, and lotion were evenly applied as single doses to human epidermal membranes mounted in static diffusion cells (not occluded). Exposures were 1-3 min (rinse-off) or 24 h (leave-on). Epidermal penetration of malic acid and lactic acid from the rinse-off shampoo and conditioner, respectively, was negligible, with >99% removed by rinsing, a negligible portion remaining in the stratum corneum (≤0.15%), and even less penetrating into the viable epidermis (≤0.04%). Glycolic acid penetration from the leave-on reference lotion was 1.42 μg equiv./cm2/h, with total absorbable dose recovery (receptor fluid plus epidermis) of 2.51%, compared to 0.009%, 0.003%, and 0.04% for the rinse-off reference lotion, shampoo (malic acid), and conditioner (lactic acid) exposures, respectively. Dermal penetration of AHAs into human skin is pH-, concentration-, and time-dependent. Alpha-hydroxy acids in rinse-off shampoos and conditioners are almost entirely removed from the skin within minutes by rinsing (resulting in negligible epidermal penetration). This suggests that ultraviolet radiation-induced skin effects of AHA-containing rinse-off products are negligible.

This paper presents the original seismic design and seismic performance retrofit study against large-scale earthquakes of the Akashi Kaikyo Bridge. The Akashi Kaikyo Bridge is the world's longest suspension bridge and was completed in 1998. In the original seismic design, an inland near-field earthquake with large magnitude was not considered for the seismic design force. Although the bridge encountered the Hyogo-ken Nanbu Earthquake in 1995, it was not seriously damaged since the earthquake occurred during its construction. Therefore, a seismic performance retrofit study against large-scale earthquakes has been started. In the seismic performance retrofit study, site-specific large-scale earthquakes including inland near-field earthquakes were defined based on the latest seismological information. As a result, it was found that target seismic performance was ensured by taking some minor countermeasures.

Transcatheter arterial chemoembolisation for hepatocellular carcinoma is widely carried out not only through the hepatic artery but also through the extrahepatic collateral pathways. Anatomically, there are many anastomoses between the hepatic artery and the extrahepatic collateral as well as among the extrahepatic collaterals. However, these anastomoses may not be shown on angiography because the anastomosing branches are too small. These anastomoses may not only interfere with effective control of hepatocellular carcinoma by transcatheter arterial chemoembolisation but also cause unexpected procedure-related complications. Therefore, radiologists should have sufficient knowledge of these underlying anastomoses. In this report, we present our angiographic images.

We developed a detection method for complete connection of a particular waterproof soft-shell electronic connector with a sliding locking device for automobiles and demonstrated a practical automatic detection system for complete connection. The proposed detection method is based on a click generated from the sliding-locking device at complete locking. However, the click generated by this type of connector is so quiet that it must be detected by a miniature microphone, amplified, and subjected to various electrical processes. Complete connection is then automatically determined from the signal level and the settling time and frequency components of the click. As a result of the demonstration of a practical automatic detection system, the detection system detected complete connection at a probability of 100% for 100 connectors and successfully rejected incomplete connections. There are limitations to the proposed detection method. The detection system did not prevent misdetection due to other types of connectors and noises that have sound components similar to those of the click for the waterproof soft-shell connector with a sliding-locking device. However, we do not consider these misdetections to be a serious problem, because sound components similar to the click for the waterproof soft-shell connector with a sliding-locking device are rare in the manufacturing environment.

It was the aim of this trial to study the effect of intranasal beclomethasone dipropionate on symptoms and signs of perennial allergic rhinitis in Japanese patients. In a multicenter trial 183 patients, children and adults, were treated with placebo or with 400 microgram beclomethasone dipropionate a day for 2 weeks. The active treatment had an effect on all nasal symptoms: sneezing, nose blowing, and blockage being reduced to 34%, 44% and 63% (P < 0.01) of the values in the placebo group. A considerable carry-over effect was found, suggesting a group comparative design to be preferable for a cross over trial for the study of intranasal steroids. The beclomethasone dipropionate therapy had a significant inhibitory effect on the immediate response to nasal allergen provocation. The number of secretion eosinophils was reduced during treatment, and the appearance of the mucous membrane tended to normalize. Local side effects were few and insignificant. It is concluded that beclomethasone dipropionate is a valuable drug for the treatment of perennial allergic rhinitis.

遠隔地間会議システム等の応用を想定して, 異なる可視化要件を持つ各種の表示オブジェクトを分散処理によりオブジェクト毎に独立に描画， これを重畳・立体視表示する手法を提案し， 実験システムにより機能・性能面で検証を行なった結果を報告する． 本手法では各オブジェクトが独立な可視化速度を持つ事が可能で， 遠隔地間会議システムにおいて高い可視化速度が要求される人物像と, 高速な可視化を要求しない他のオブジェクトを同時に表示する場合に有効である． 提案手法を実装した実験システムにより, 異なる可視化要件を持つ表示オブジェクトの仮想会議室空間への配置と立体視表示， 複数オブジェクトの同時表示機能を確認した. 性能面では， 複数オブジェクト同時表示時に相互の可視化速度へ与える影響が少ない事を確認し, 人物像の高速な可視化には, 描画処理と重畳処理間のデータ転送の高速化が必要である事を確認した.

We have developed a solder joint material with a plastic core. In this paper, we report on the thermal conductivity of this material. We evaluated the thermal resistance /spl theta/ j-a with 352pin EBGA and 352pin PBGA, and also carried out a thermal conductivity analysis using computer simulation. The result shows that the /spl theta/ j-a of the 352pin EBGA was 12/spl deg/C/W, /spl theta/ j-a of the 352pin PBGA was 20/spl deg/C/W. So this material is applicable for high frequency and power package (CPU, MCM, MCP, stacked package and so on).

A hybrid-emulsion experiment has been carried out at KEK-PS to study S = −2 nuclei. With the analysis of about 35% of total emulsion, we have successfully found three events of sequential weak decay of a double-hypernucleus. The results of these E373 events are summarized together with those of past experiments. The first experimental study of the branching ratios of weak decay modes of double-hypernuclei has been performed.

It has been known that dendritic cells (DCs) including Langerhans cells (LCs) play a critical role in the skin sensitization process. Many attempts have been made to develop in vitro sensitization tests that employ DCs derived from peripheral blood mononuclear cells (PBMC-DC) or CD34+ hematopoietic progenitor cells (CD34+ HPC) purified from cord blood or bone marrow. However, the use of the DCs in in vitro methods has been difficult due to the nature of these cells such as low levels in the source and/or donor-to-donor variability. In our studies, we employed the human monocytic leukemia cell line, THP-1, in order to avoid some of these difficulties. At the start, we examined whether treatment of the cells with various cytokines could produce DCs from THP-1. Treatment of THP-1 cells with cytokines such as GM-CSF, IL-4, TNF-alpha, and/or PMA did induce some phenotypic changes in THP-1 cells that were characteristic of DCs. Subsequently, responses to a known sensitizer, dinitrochlorobenzene (DNCB), and a non-sensitizer, dimethyl sulfoxide (DMSO) or sodium lauryl sulfate (SLS), on the expression of co-stimulatory molecules, CD54 and CD86, were examined between the naive cells and the cytokine-treated cells. Interestingly, the naive THP-1 cells responded only to DNCB and the response to the sensitizer was more distinct than cytokine-treated THP-1 cells. Similar phenomena were also observed in the human myeloid leukemia cell line, KG-1. Furthermore, with treatment of DNCB, naive THP-1 cells showed augmented expression of HLA, CD80 and secretion of IL-1 beta. The response of THP-1 cells to a sensitizer was similar to that of LCs/DCs. Upon demonstrating the differentiation of monocyte cells in our system, we then evaluated a series of chemicals, including known sensitizers and non-sensitizers, for their potential to augment CD54 and CD86 expression on naive THP-1 cells. Indeed, known sensitizers such as PPD and 2-MBT significantly augmented CD54 and CD86 expression in a dose-dependent manner while non-sensitizers, such as SLS and methyl salicylate (MS), did not. To note, the metal allergens such as (NH(4))(2)[PtCl(4)], NiSO(4) and CoSO(4) augmented significantly only CD54 expression. Taking advantage of a cultured cell line, measurement of the co-stimulatory molecules, CD54 and CD86, on naive THP-1 cells following chemical exposure shows promise for the development of a simple, short-term in vitro sensitization test.

Melon yellow spot virus (MYSV) and Watermelon silver mottle virus (WSMoV) are distinct virus species belonging to the genus Tospovirus in the Family Bunyaviridae. They have quasi-spherical virions with lipid envelopes where three single-stranded RNA segments are contained, and are persistently transmitted by Thrips palmi. WSMoV was first identified in 1982, and it has been causing problems on watermelon, wax gourd, melon, cucumber and bitter gourd productions in Japan and Taiwan. MYSV was first identified in 1992, and it has been causing severe damages on melon and cucumber productions in Japan. Nucleotide sequences of the smallest RNA segments of WSMoV and MYSV were analyzed to examine genetic relationships. Identities of the nucleocapsid protein genes of WSMoV or MYSV with other tospovirus species suggested that MYSV and Physalis severe mottle virus (PYSV) were considerably similar. Identities of the nucleocapsid protein genes and the non-structural genes of two isolates of MYSV from cucumber plants with those of reported MYSV with those with PSMV were more than 96%. Therefore, PSMV and MYSV should be classified as the same species. Comparison of Japanese isolates and Taiwanese isolates of WSMoV suggested that they are genetically related each other, except for a 20 nucleotide insertion sequence was found in Japanese isolates. RT-PCR assay with specific primers of WSMoV or MYSV were examined. RT-PCR with WSMoV specific primers WN5 (5' -GCACACACATAGGTATAACT-3') and WN3 (5' -TCCTTTGTAAACACCATG-3') detected approximately 1.1 kb DNA fragment from WSMoV-infected samples. RT-PCR with MYSV specific primers MN5 (5' TATTTCATTCAACTAGTTAA3') and MN3 (5' ATTCAACATCAGCAAGTCAA3') detected approximately 1.0 kb DNA fragment only from MYSV-infected samples. No significant amplification was detected from other tospovirus -infected samples or healthy samples.

Objective The aim of this observational study was to construct an ultrasound index of fetal gastric size for the prenatal detection of congenital digestive tract obstruction. Subjects A total of 386 fetal measurements were performed in routine ultrasonographic examinations of women with normal singleton pregnancies between 18 and 39 weeks of gestation. Gastric measurements were also performed in 13 fetuses with digestive tract obstruction. Methods The ultrasound plane which included the pylorus and which provided the largest stomach area was used for definition and measurement of gastric area and maximal longitudinal dimension. The transverse section at the center of the gastric corpus was used for transverse and anteroposterior dimensions. Gastric volumes were calculated as a prolate ellipsoid. The gastric area ratio was defined as the gastric area divided by the transverse abdominal area. Biparietal diameter (BPD) and abdominal transverse area were also measured. Results The fetal gastric area was significantly correlated with fetal gastric volume (r = 0.91) and gestational age (r = 0.74). However, the correlation coefficient for gastric area with gestational age was smaller than those of the BPD (r = 0.97) with gestational age and abdominal transverse area with gestational age (r = 0.97). Gastric area ratio decreased slightly towards term. The gastric area ratio was below the 95% confidence intervals for the predicted values in all five fetuses with esophageal atresia, and exceeded the 95% confidence intervals in seven of the eight fetuses with duodenal atresia or intestinal tract obstruction. Conclusion Fetal gastric area correlates with ultrasound-determined gastric volume measurements and appears to be useful in the assessment of digestive tract anomalies. Copyright © 2002 ISUOG

We have focused on the magnetic memories using parallel-aligned nanowires without mechanical moving parts, in order to achieve the ultra-high transfer rate of more than 72 Gbps for future ultra-high definition TV. It is important for the storage devices, where the magnetic nanowires are utilized, that the trapping energy of the magnetic domains should be optimized precisely. In order to tune this energy finely, we have fabricated ultra-small notches less than 20 nm in the z-direction of [Co/Pd] nanowires with perpendicular magnetic anisotropy by the nano-scratch, anodic oxidation and nano-indentation method, instead of the conventional notches fabricated in-plane of nanowires. We have succeeded to entrap magnetic domain walls by these trap sites on the nanowire.

The cyclin-dependent kinase (CDK) inhibitor, flavopiridol, was tested as a potential new cancer therapeutic agent to treat canine lymphoma by examining its effect on cell growth of canine lymphoma cell lines in vitro. Flavopiridol induced profound cell death in all eight lymphoma cell lines at 400 nM, and in all cases cell death was due to apoptosis. Apoptosis was inhibited by caspase inhibitor, despite the variable sensitivities between cell lines. Analysis of the mechanism of flavopiridol-induced apoptosis showed that Rb phosphorylation was inhibited, possibly due to CDK4 or CDK6 inhibition. There was also decreased expression of Rb protein and anti-apoptotic proteins, Mcl-1 and XIAP, possibly through transcriptional regulation by inhibition of CDK7 or CDK9 activation. Canine lymphoma cell line-xenotransplanted mice were then treated with flavopiridol and profound tumour shrinkage was observed. This study describes a new therapeutic approach using flavopiridol for canine lymphoma treatment.

Reovirus is a potent oncolytic virus in many human neoplasms that has reached phase II and III clinical trials. Our laboratory has previously reported the oncolytic effects of reovirus in canine mast cell tumour (MCT). In order to further explore the potential of reovirus in veterinary oncology, we tested the susceptibility of reovirus in 10 canine lymphoma cell lines. Reovirus-induced cell death, virus replication and infectivity were confirmed in four cell lines with variable levels of susceptibility. The level of Ras activation varied among the cell lines with no correlation with reovirus susceptibility. Reovirus-susceptible cell lines underwent apoptosis as proven by propidium iodide (PI) staining, Annexin V-FITC/PI assay, cleavage of PARP and inhibition of cell death by caspase inhibitor. A single intratumoral injection of reovirus suppressed the growth of canine lymphoma subcutaneous tumour in NOD/SCID mice. Unlike canine MCT, canine lymphoma is less susceptible to reovirus.