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The deposition behavior of sprayed individual metallic particles on the substrate surface in the cold spray process was fundamentally investigated. As a preliminary experiment, pure copper (Cu) particles were sprayed on mirror-polished stainless steel and aluminum (Al) alloy substrate surfaces. Process parameters that changed systematically were particle diameter, working gas, gas pressure, gas temperature, and substrate temperature, and the effect of these parameters on the flattening or adhesive behavior of an individual particle was precisely investigated. Deposition ratio on the substrate surface was also evaluated using these parameters. From the results obtained, it was quite noticeable that the higher substrate temperature brought about a higher deposition rate of Cu particles, even under the condition where particles were kept at room temperature. This tendency was promoted more effectively using helium instead of air or nitrogen as a working gas. Both higher velocity and temperature of the particles sprayed are the necessary conditions for the higher deposition ratio in the cold spraying. However, instead of particle heating, substrate heating may bring about the equivalent effect for particle deposition.

Background Enhanced expression of CD14+CD15+ cells has been reported in the bone marrow of patient with severe rheumatoid arthritis (RA) (Tomita et al, J. Rheumatology, 1997). CD14+CD15+ cells (the phenotype of previously described “abnormal” myeloid cells) has been also reported to be increased in peripheral blood stem cell harvests on a per CD34+cell basis (Snowden et al, Br. J of Haematol, 1998) suggesting the abnormal process in the early differentiation of bone marrow cells in RA patients. Our study is focusing to clarify the appearance of CD14+CD15+ population in the early differentiation of bone marrow cells from RA patients. As the strategy, we utilize the monocyte differentiation system of induced pluripotent stem (iPS) cells derived from RA patients. Objectives To examine whether the CD14+CD15+ cells appear during hematologic differentiation process of the iPS cells established from RA patients (RA-iPS). Methods iPS cells were established from skin fibroblasts and peripheral blood mononuclear cells from RA patients as described previously. Six days after transduction, the cells were harvested and re-plated onto mitotically inactivated SNL feeder cells. For the healthy controls, non-onset family members (NOF) of the patients are recruited as donors in order to adjust hereditary background as much as possible. Monocytes were induced from each iPS clone culture on feeder-free conditions using a previously reported method (Niwa et al, PLoS One, 2011). The cells collected from each culture were stained with specified antibodies against CDs, including CD14 and CD15 and were analyzed by fluorescence-activated cell sorting (FACS) on day 12, 15, 18, 21 and 25. Results Cultured cells started to express CD14 on the day 12 and more than 90% of the cells expressed CD14 on the day 21 in the monocyte differentiation induction course. According to the expression levels of CD14, the cell population was divided into three groups: CD14 (−), CD14 (+) and CD14 (++). CD15 (+) cells were observed in CD14 (−) and CD14 (+) population but not in CD14 (++) population. The CD15+ cells in CD14 (+) transiently appeared in RA-iPS derived cells at 11.9±2.8% (mean ± SE) on day15. However these cell proportion in NOF was1.7±2.0%. Meanwhile, CD15+ cells in CD14 (−) proportion decreased during monocyte differentiation in RA-iPS cells, but remained in NOF-iPS cells (representative data, RA 31.5, 20.6, 15.6%, NOF 47.3, 46.1, 47.3%, on day15, 18 and 21). Conclusions We successfully differentiated iPS cells into monocytes and found CD14+CD15+cells transiently appeared only in the very early differentiation process of RA-iPS cells into monocytes. Taken together with the evidence that CD14+CD15+cells exist in RA patient bone marrow, abnormal early differentiation of myeloid/monocyte lineage cells may be pathologically involved in RA. Functional analysis will be required to elucidate the role of this distinct CD14+CD15+population in RA. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.3073

BackgroundRheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic proliferation of synovial cells and destructive polyarthritis. Bone destruction in RA is mediated by increased numbers of osteoclasts, and the enhanced osteoclastgenesis has been reported in RA. However, it remains unclear whether or not the genetic factors influence the increased osteoclastgenesis in RA. To answer this question, we induced osteoclasts in vitro using induced pluripotent stem cells (iPSC) derived from RA-patients and non-onset family members of the patients (NOR).ObjectivesThe aim of this study is to clarify if the osteoclastgenesis in RA is enhanced genetically compare to NOR.MethodsiPSCs have been established from skin fibroblasts from RA patients and NORs. Under monocytes-induced conditions in vitro, floating cells were sequentially collected and subsequently inoculated onto glass chamber slides with M-CSF and RANKL. An osteoclast was defined as a TRAP-positive cell containing at least three nuclei. To analyze the bone resorption activity, the fluorescent-intensity of FITC-labeled chondroitin sulfate (CS), which is released from the calcium phosphate layer resolved by osteoclasts, was measured in the culture supernatant by a fluorimeter. The expressions of osteoclast markers (cathepsin K and MMP-9) in osteoclasts induced from monocytes collected on day 24 and day 32, and osteoclast-precursor markers (CD115: MCSF-receptor, and RANK) in the monocytes collected on days 24, 28 and 32 after induction from iPSCs were analyzed by either RT-PCR or fluorescence-activated cell sorting using the corresponding antibodies.ResultsOsteoclastgenesis of monocytes derived from RA-iPSCs increased as compared to that of NOR-iPSCs. The TRAP positive cell numbers per well of RA patients were higher than those of NOR in the presence of RANKL (25 ng/mL and 100 ng/mL) at all 6 time points as shown in the figure below. The fluorescent-intensity of FITC-labeled CS in the culture supernatant of osteoclasts differentiated from RA-iPSCs was also higher than that of NOR-iPSCs (fluorescent intensity/well; 4869±838 and 954±252, respectively on day24). The expression of cathepsin K and MMP9 on day24 were also increased in osteoclasts from RA-iPSCs by 20 and 12 times higher than osteoclasts from NOR-iPSCs, respectively. The proportion of CD115-positive cells in CD14-positive cells was increased in monocytes induced from RA-iPSCs. Furthermore, monocytes induced from RA-iPSCs highly expressed RANK mRNA as compared with those of NOR-iPSCs at all day points of harvesting monocytes.ConclusionsThe genetically up-regulated M-CSF receptor and RANK in the osteoclast precursors in RA-iPSC may lead to the enhanced osteoclastgenesis.Disclosure of InterestNone declared

Ion-exchange treatment is a promising technique for removing hydrophilic compounds during drinking water treatment. In this study, we applied several different ion exchangers (i.e., anion exchange resins and a hydrotalcite compound) to bromide removal to minimize bromate formation during ozonation. It was found that ion-exchange treatment affected ozone and hydroxyl radical concentration profiles as well as bromate ion concentration after ozonation. Selecting an appropriate ion exchanger is important to achieve both the oxidation of target contaminants and the reduction of bromate ion during ozonation.

Chronic myelomonocytic leukemia (CMML) is an entity of myelodysplastic syndrome/myeloproliferative neoplasm. Although CMML can be cured with allogeneic stem cell transplantation, its prognosis is generally very poor due to the limited efficacy of chemotherapy and to the patient’s age, which is usually not eligible for transplantation. Comprehensive analysis of CMML pathophysiology and the development of therapeutic agents have been limited partly due to the lack of cell lines in CMML and the limited developments of mouse models. After successfully establishing patient’s derived disease-specific induced pluripotent stem cells (iPSCs) derived from a patient with CMML, we utilized these CMML-iPSCs to achieve hematopoietic re-differentiation in vitro , created a humanized CMML mouse model via teratomas, and developed a drug-testing system. The clinical characteristics of CMML were recapitulated following hematopoietic re-differentiation in vitro and a humanized CMML mouse model in vivo . The drug-testing system using CMML-iPSCs identified a MEK inhibitor, a Ras inhibitor, and liposomal clodronate as potential drugs for treating CMML. Clodronate is a drug commonly used as a bisphosphonate for osteoporosis. In this study, the liposomalization of clodronate enhanced its effectiveness in these assays, suggesting that this variation of clodronate may be adopted as a repositioned drug for CMML therapy.

Quantitative real-time PCR shows the quantity in addition to the presence of the target sequence. This property seemed very useful for library screening. Then, real-time PCR was employed to screen for lambda phages carrying D-amino-acid oxidase gene from mouse genomic library. Using stepwise dilution screening combined with real-time PCR, positive phages were isolated in a short time.

The Rescue PT system consists of a flexible, dual lumen Monorail 4.5 French catheter and a collection bottle with a filter for separating solid bodies from blood that contains small pieces of thrombus and atheromatous plaque. Coronary artery angiography showed the defects decreased after aspiration therapy. [...]PCI was easily successful with adjunctive stenting.

Previous studies showed that the cellular amino acid composition obtained by amino acid analysis of whole cells, differs such as eubacteria, protozoa, fungi and mammalian cells. These results suggest that the difference in the cellular amino acid composition reflects biological changes as the result of evolution. However, the basic pattern of cellular amino acid composition was relatively constant in all organisms examined. In the present study, we examined archaeobacteria, because they are considered important in understanding the relationship between biological evolution and cellular amino acid composition. The cellular amino acid compositions of Archaeoglobus fulgidus, Pyrococcus horikoshii, Methanobacterium thermoautotrophicum and Methanococcus jannaschii differed slightly from each other, but were similar to those determined from codon usage data, based on the complete genomes. Thus, the cellular amino acid composition reflects biological evolution. We suggest that primitive forms of life appearing on earth at the end of prebiotic evolution had a similar-cellular amino acid composition.

The cellular amino acid composition of plant cells was analyzed. The callus of carrot (Daucus carota), leaves of Torenia fournieri and protocomb-like body of Cymbidium, s.p. were examined as examples of plant cells. The cellular amino acid compositions differed in the plant cells, but their basic patterns were quite similar. It is concluded that the basic pattern of the cellular amino acid composition is conserved in all terrestrial organisms, including plants.

The number of hospitals in Japan exceeds 10,000, and every month nurses are scheduled to shifts in about 30,000 units in total. There is serious demand for automating this scheduling task. In this paper, we introduce a mathematical programming formulation of the nurse scheduling problem in Japan, and develop a meta-heuristic approach to solve the problem. This scheduling problem is a hard combinatorial problem due to tight constraints involving such factors as the skill level of a team, the need to balance workload among nurses, and the consideration of nurses' preferences, even though the number of the nurses to be scheduled is not large, at between 20 and 40. The performance of our approach is demonstrated by the successful solution of data taken from actual scheduling problems. The proposed model and approach can be adapted for the majority of hospitals in Japan, as well as for some hospitals in other countries, and is likely applicable to many other scheduling problems in the fields of business and logistics.