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The aim of this study was to clarify the clinic-level factors related to experiences of and attitudes toward collaboration with community pharmacies. We conducted a postal questionnaire survey of all clinics in Gifu, Japan, assessing the experiences and attitudes of representative clinical staff regarding the following activities in collaboration with community pharmacists: regional care meetings/service adjustment meetings, case study conferences, joint workshops/continuing education conferences, community services, information sharing through medical cooperation networks, and accompanying community pharmacists during home care. The factors significantly related to experiences of joint workshops/continuing education conferences included home care visits (odds ratio [OR] 2.39) and a 100 % out-of-hospital prescription ratio (OR 4.80). In contrast, only home care visits were significantly associated with consideration of information sharing through medical cooperation networks and accompanying community pharmacists during home care (OR 2.06 and 11.91, respectively). Finally, the factors significantly associated with considering implementing case study conferences and joint workshops/continuing education conferences included home care visits (OR 4.64 and 2.98, respectively) and a 100% out-of-hospital prescription ratio (OR 4.64 and 6.38). Overall, having more opportunities to communicate with community pharmacists and other healthcare professionals appeared to facilitate clinics' consideration of collaboration with community pharmacies, along with actual experiences.

Water striders are insects living on the water surface, over which they can move very quickly and rarely get wetted. We measured the force of free walking in water striders, using a hair attached to their backs and a 3D strain gauge. The error was calculated by comparing force and data derived from geometry and was estimated as 13%. Females on average were stronger (1.32 mN) than males (0.87 mN), however, the ratio of force to weight was not significantly different. Compared with other lighter species, Aquarius paludumseems stronger, but the ratio of force to weight is actually lower. A. paludumapplies about 0.3 mN·cm−1to 0.4 mN·cm−1with its mid-legs, thus avoiding penetrating the surface tension layer while propelling itself rapidly over the water surface. We also investigated the external morphology with SEM. The body is covered by effectively two layers of macro-and micro-hairs, which renders them hydrophobic. The setae are long (40 µm–60 µm) and stiff, being responsible for waterproofing, and the microtrichia are much smaller (<10 µm), slender, and flexible, holding a bubble over the body when submerged.

Spark assisted chemical engraving (SACE) is a method for 3D microstructuring of glass or other non-conductive materials with high aspect ratio and smooth surface quality. It is applicable for rapid prototyping of microfluidic devices, for MEMS interfacing and similar applications. Typical feature size is in the hundreds of micrometres, down to a few tens of micrometres. It is a table-top technology requiring no clean rooms and no masks and with very modest space usage. It is thus well suited for microfactories. This paper gives a basic introduction to SACE and some machining examples.

Spark assisted chemical engraving (SACE) is an unconventional micromachining technology based on electrochemical discharge phenomena for glass and various ceramics. The limits of SACE with respect to small dimensions in the particular case of glass are explored. It is found, using a specially developed set-up based on an AFM, that even using extremely sharp tool-electrodes does not allow us to produce a smaller pattern than typically 25 µm. It is concluded that the gas film thickness, in which the electrochemical discharges take place, is the main limiting factor. Further experimental investigations on its formation are investigated. By adding surfactants to the electrolyte, in order to increase the wettability of the tool-electrode and therefore to reduce the gas film thickness, it is observed experimentally that the critical voltage reduces significantly. This observation may lead to a novel method of characterizing the gas film thickness in SACE.

Murine monoclonal antibodies (mAbs) against Neospora caninum tachyzoites were produced to identify the cross-reactive antigens between N. caninum and Toxoplasma gondii. Ten mAbs recognizing cross-reactive antigens of both parasites were obtained and tentatively classified into 6 different groups based on their reactivity patterns in an indirect fluorescent antibody test and Western blot analysis. Three mAbs in group 1 recognized antigens located on the surface of parasites with molecular masses ranging from 28 to 76 kDa; one mAb in group 2 recognized antigens located on interior organelles of parasites with a molecular mass of 50 kDa; one mAb in group 3 recognized antigens located on interior organelles of parasites with molecular masses of 35 kDa and 14 kDa; three mAbs in group 4 recognized antigens located on interior organelles with a molecular mass of 64 kDa; one mAb in group 5 recognized antigens located on the surface of parasites with an unknown molecular mass; one mAb in group 6 recognized antigens located on the apical end of parasites with an unknown molecular mass. The mAbs in groups 1, 2, 3, and 5 showed inhibitory effects on the growth of the two parasites in vitro in a dose-dependent manner. A cDNA expression library prepared from N. caninum tachyzoite mRNA was immunoscreened with the mAb panel. Three kinds of proteins, protein disulfide isomerase (PDI), heat-shock protein 70 (HSP70), and ribosomal protein 1 (RP1), were identified as cross-reactive antigens recognized by mAbs in groups 2, 3, and 4, respectively. Some of the proteins could be useful in developing vaccines or drugs for controlling the diseases caused by the two parasites.

We evaluated the growth inhibitory efficacy of the imidazole derivatives, clotrimazole (CLT) and ketoconazole (KC), and the herbicide clodinafop-propargyl (CP), in in vitro cultures of Babesia bovis and B. bigemina. Clotrimazole was effective in a dose range of 15 to 60 μM (IC50: 11 and 23·5 μM), followed by KC (50 to 100 μM; IC50: 50 and 32 μM) and CP (500 μM; IC50: 265 and 390 μM). In transmission electron microscopy, extensive damage was observed in the cytoplasm of drug-treated parasites. Combinations of CLT/KC, CLT/CP and CLT/KC/CP acted synergistically in both parasites. In contrast, the combination of KC/CP was exclusively effective in B. bovis, but not in B. bigemina.

In the present study, morphological observations on the extracellular structures found on the apical surface of the midgut epithelium, known as the peritrophic membrane (PM) or glycocalyx, are described inHaemaphysalis longicornisfemales and larvae. These structures have been hypothesized to provide protection to the microvilli of epithelial cells of the digestive tract. Our aim was to determine whether the extracellular structures are important in the digestion of the blood meal and/or as a protection against infection or injury. The PM was detectable in the midgut of engorged larvae by electron microscopy, but not in engorged females. However, a PM-like structure, stainable with toluidine blue, was observed in females by light microscopy. From the results of confocal laser scanning and electron microscopic observations with wheat germ agglutinin (WGA lectin) staining for chitin of the PM, however, the structure was clearly recognized. The structure in the female is likely to be PM because staining with WGA lectin in the presence of GlcNAc indicates the presence of chitin and various morphologies of PM have been reported in insects and ticks. These results show morphologically that different types of PM-like structure are formed in larvae and females ofH. longicornis.

The initial solidification in the continuous casting process can be controlled by imposing a pulsative AC electromagnetic field on the meniscus. The casting experiment of a steel billet has been conducted by using a commercial billet caster. The imposition of a pulsative AC electromagnetic field has greatly improved the surface qualities. This presentation provides the results of the electromagnetic casting project of JRCM which is supported financially by the Ministry of International Trade and Industry of Japan.

The initial solidification in the continuous casting process can be controlled by imposing a pulsative AC electromagnetic field on the meniscus. The casting experiment of a steel billet has been conducted by using a commercial billet caster. The imposition of a pulsative AC electromagnetic field has greatly improved the surface qualities. This presentation provides the results of the electromagnetic casting project of JRCM which is supported financially by the Ministry of International Trade and Industry of Japan.

Interferon-gamma (IFN-γ) has a crucial role for host defence against parasite infection. It is not clear, however, how IFN-γ affects the parasite-infected host cells. The effect of IFN-γ on Neospora caninum-infected cells was investigated in murine fibroblasts and canine kidney cells in vitro. In the presence of IFN-γ, the viability of the infected host cell was decreased and apoptotic cell death occurred, as analysed by DNA stainings with propidium iodide and a terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labelling (TUNEL) and DNA fragmentation. The percentage of apoptotic cells depended on the dose of IFN-γ. Flow cytometric analysis indicated a significant increase of FasL expression on the IFN-γ-treated cells following N. caninum infection. Moreover, IFN-γ treatment down-regulated Bcl-2 expression in the cells cultured with N. caninum while parasite infection up-regulated Bcl-2 expression. The present study suggests that the IFN-γ induced increases of FasL expression and down-regulated Bcl-2 expression in N. caninum-infected cells are associated with apoptosis in vitro.

Abstract: We describe the expression of SAG-1 cDNA in B6C3F1 mice by microinjecting a 3.3 kbp DNA fragment, consisting of the cytomegalovirus enhancer-chicken β-actin hybrid promoter and SAG-1 into the pronucleus of a fertilized egg at the one-cell stage. Offspring derived from this microinjection were analyzed for the integration and functional expression of the SAG-1 transgene. Steady-state expressions of both the mRNA for SAG-1 and SAG-1 protein product were detected in the brain, thymus, spleen and liver. Approximately 50% of F1 and F2 progeny inherited the SAG-1 transgene from SAG-1 transgenic mice in Mendelian fashion. These results indicated that SAG-1 transgenic lines were established. Transgenic mice harboring the SAG-1 gene will contribute a critical tool of defining the molecular mechanisms of SAG-1 in pathogenesis and host immune response.

Serological comparisons among piroplasm antigens of the benignTheileria species of cattle from Japan, Australia and Britain, which are frequently referred to asT. sergenti, T. buffeli andT. orientalis, were carried out. The results obtained from comparative enzyme-linked immunosorbent assay (ELISA) using sera from infected cattle suggest thatT. sergenti could be differentiated from bothT. buffeli andT. orientalis by their serological dissimilarities. Western blotting combined with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) revealed that both the 33-kDa piroplasm protein ofT. sergenti and the similar 34-kDa protein ofT. buffeli andT. orientalis corresponded to immunodominant antigens against cattle. The other 32-kDa proteins ofT. buffeli andT. orientalis also represented immunodominant antigens. Cross-reactivities of the 32-and 34-kDa proteins were observed betweenT. buffeli andT. orientalis, whereas the 33-kDa protein ofT. sergenti could be differentiated from the similar 34-kDa proteins ofT. buffeli/orientalis. The present study suggests thatT. sergenti should be separated fromT. buffeli andT. orientalis on the basis of their serological dissimilarities.

Benign Theileria species in cattle, Theileria sergenti and T. buffeli, are morphologically indistinguishable. The polymerase chain reaction (PCR) was used to amplify the genes encoding the 33- and 34-kDa major piroplasm antigens (p33/34) of T. sergenti and T. buffeli from cattle blood infected with these parasites and tick salivary gland infected with T. sergenti. Following amplification, the p33 gene from T. sergenti and the p34 gene from T. buffeli were clearly differentiated using the restriction enzyme sites that were not shared between them. The oligonucleotide primer set, designed from the p33/34 genes, was specific for these Theileria species, since no amplification was detected with DNA from Babesia ovata, B. bovis, Anaplasma marginale, A. centrale, Eperythrozoon wenyoni, bovine white blood cells, and uninfected tick salivary glands. One tenth vol of the template prepared from either 25 µl of blood with 0.5% parasitemia or individual tick salivary glands with six infected acini allowed sufficient amplification for differentiation of the two parasite species by restriction enzyme digestion. In addition, this system could be used to demonstrate the simultaneous, experimentally induced infection of cattle with T. sergenti and T. buffeli. The PCR-based marker system therefore provides a means to differentiate T. sergenti from T. buffeli in infected cattle blood and infected tick salivary glands. This system may also be useful for the characterization of other benign Theileria species in cattle.Copyright 1995, 1999 Academic Press, Inc.

A method of obtaining a pure suspension of Theileria sergentipiroplasms from infected bovine erythrocytes was developed and the resulting parasites used as antigen in an enzyme-linked immunosorbent assay (elisa). Piroplasms were freed from infected erythrocytes using the nitrogen cavitation technique and then separated from unbroken erythrocytes by centrifugation at 670 gmax. The parasites in the supernatant were then sedimented by centrifugation at 2700 gmaxand the purified fraction examined by electron microscopy. This examination showed that the isolated piroplasms were morphologically intact and that there were few contaminants. sds-pageand spectrophotometric analysis showed that this fraction contained little erythrocyte component. The piroplasms thus obtained were sonicated and treated with Triton X-100 then used for elisaantigen. The elisavalues had a high correlation with titres obtained using the indirect fluorescent antibody test on sera from cattle infected with T sergenti.

This paper presents a spatial filtering method for reducing ill-effects of atmospheric turbulence on holographic imaging using airborne radars. The method is based on the spatially weighting of amplitudes of received waves and is examined by showing graphically the average strip-image which is holographically reconstructed by using exponential weight functions. The introduction of the weight functions yields fairly good restoration of images degraded by atmospheric turbulence and may be useful for image reconstruction in the case of extremely strong turbulence.

A simple and rapid method for purifying merozoites ofTheileria sergenti from infected bovine erythrocytes was developed. Infected erythrocytes were lysed by the use of a cytolytic toxin produced byAeromonas hydrophila and the lysate was subjected to ultracentrifugation in a Percoll discontinuous density gradient. Pure and morphologically intact merozoites free of erythrocyte membrane were recovered from a band formed at the interface of 40% and 60% (by vol.) Percoll solutions. In this merozoite fraction, contamination of erythrocyte membrane proteins was not detected as examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

A latex agglutination test (LAT) using recombinant equi merozoite antigen 1 (EMA-1) for the detection of antibodies to Babesia equi was developed. The LAT was able to differentiate very clearly between sera from B. equi-infected horses and sera from Babesia caballi-infected horses or from normal horses. The LAT results were identical to those of a previously developed enzyme-linked immunosorbent assay. These results indicate that LAT using recombinant EMA-1 might be very useful as a routine screening method for the diagnosis of B. equi infection.

The ultrastructure and characteristics of hemocytes of argasid tick species, Ornithodoros moubata, during the ecdysdial phase are herein presented. Hemocyte classes/populations characterized based on their affinity with Giemsa stain and ultrastructural differences comprised the prohemocytes, nongranular cells (Nc), eosinophilic granular cells (Ec), basophilic granular cells (Bc), and unidentified cells. Significant changes/shift in the ratio of hemocyte classes/population was apparent in ticks before and after the ecdysial phase. The granule-scant basophilic granular cells (sBc) constituted the most abundant hemocyte population in the ecdysial phase. Nymphs in ecdysis showed increases in Nc and sBc and decrease in Ec, a phenomenon that was reversed in unengorged nymphs and adults ticks. The significant increase in total Bc population in ecdysis relative to nonengorged ticks clearly point to blastogenesis of Bc taking place during the ecdysial phase and Bc’s important role in the process of tissue remodeling.