Ligand-competent fractalkine receptor is expressed on exosomes

Expression of chemokine receptor CX3CR1 is reportedly restricted to several cell types including natural killer cells, cytotoxic T cells, monocytes, and macrophages. However, its expression and function on exosomes, which are nanosized extracellular vesicles known to act as mediators of intercellular communications, remain unclear. Here, we investigated CX3CR1 expression on exosomes isolated from various cell types. Although we found that all the exosomes tested in our study highly expressed CX3CR1, this chemokine receptor was expressed only inside, but barely on, their source cells. Moreover, exosomal CX3CR1 was capable of binding soluble CX3CL1. Therefore, our study suggests that CX3CR1 is a novel and ligand-competent exosome receptor.     

Analysis of chemical modification of RNA from formalin-fixed samples and optimization of molecular biology applications for such samples.

Formalin-fixed archival samples are known to be poor materials for molecular biological applications. We conducted a series of experiments to understand the alterations in RNA in fixed tissue. We found that formalin-fixed tissue was resistant to solubilization by chaotropic agents. However, proteinase K completely solubilized the fixed tissue and enabled the extraction of almost the same amount of RNA as from a fresh sample. The extracted RNA did not show apparent degradation. However, as reported, successful PCR amplification was limited to short targets. The nature of such 'fixed' RNA was analyzed using synthetic homo-oligo RNAs. The heterogeneous increase in molecular weight of the RNAs, measured by MALDI-TOF mass spectrometry, showed that all four bases showed addition of mono-methylol (-CH(2)OH) groups at various rates. The modification rate varied from 40% for adenine to 4% for uracil. In addition, some adenines underwent dimerization through methylene bridging. The majority of the methylol groups, however, could be removed from bases by simply elevating the temperature in formalin-free buffer. This demodification proved effective in restoring the template activity of RNA from fixed tissue. The improvement in PCR results suggested that more than half of the modification was removed by this demodification.     

Efficient Repeated Use of Alcohol Dehydrogenase with NAD + Regeneration in an Aqueous-organic Two-phase System

Bioconversion of cinnamyl alcohol to cinnamaldehyde was carried out in an aqueous-organic two-phase reaction system by the repeated use of horse liver alcohol dehydrogenase (HLADH) and NAD + with coenzyme regeneration. Both HLADH and the coenzyme were efficiently entrapped in the aqueous phase, while the substrate was supplied successively from the organic phase and the product was accumulated in the organic phase. Optimum conditions for cinnamaldehyde production in the aqueous-organic two-phase system were also examined, including substrate concentration, pH, and organic solvent type. Under suitable conditions, both HLADH and NAD + in the aqueous-organic two-phase system could be reused, and NAD + cycling numbers of 3040 were obtained after repeated operation for 40 λh.     

Irisin supports integrin-mediated cell adhesion of lymphocytes

Irisin, a myokine released from skeletal muscle, has recently been found to act as a ligand for the integrins αVβ5, αVβ1, and α5β1 expressed on mesenchymal cells, thereby playing an important role in the metabolic remodeling of the bone, skeletal muscle and adipose tissues. Although the immune-modulatory effects of irisin in chronic inflammation have been documented, its interactions with lymphocytic integrins have yet to be elucidated. Here, we show that irisin supports the cell adhesion of human and mouse lymphocytes. Cell adhesion assays using a panel of inhibitory antibodies to integrins have shown that irisin-mediated lymphocyte adhesion involves multiple integrins including not only α4β1 and α5β1, but also leukocyte-specific αLβ2 and α4β7. Importantly, mouse lymphocytic TK-1 cells that lack the expression of β1 integrins have exhibited αLβ2- and α4β7-mediated cell adhesion to irisin. Irisin has also been demonstrated to bind to purified recombinant integrin αLβ2 and α4β7 proteins. Thus, irisin represents a novel ligand for integrin αLβ2 and α4β7, capable of supporting lymphocyte cell adhesion independently of β1 integrins. These results suggest that irisin may play an important role in regulating lymphocyte adhesion and migration in the inflamed vasculature.     

Dynamics and distribution patterns of acarine populations in stored-oat ecosystems

Population dynamics and aggregation patterns of nine kinds of stored-grain mites were studied in two 7.5 tonne lots of hulled (cv. Random) and hulless (cv. Terra) oat cultivars with 12–14% moisture content stored in two wooden bins in Manitoba, Canada during 1978–84. Random oats harbored more mites than Terra oats. Lepidoglyphus destructor Schrank was the most common granivorous mite and Cheyletus eruditus Shrank the most common predatory mite. Ecological data on a tydeid mite, Paratriophtydeus coineaui André, are presented for the first time. Because they were more abundant in Terra oats with higher fat acidity values (FAV) than Random oats with lower FAVs, L. destructor and Tarsonemus granarius Lindquist could be used as bioindicators of spoilage of stored oats. All species analyzed showed some significant difference in their abundance at different depths in the grain bulk; some species showed depthxtime interaction. Aggregation patterns indicated most mite species had overdispersed (clumped) distribution. T. granarius, and Blattisocius keegani Fox-Androlaelaps casalis Berlese had a distinct aggregation pattern in each oat cultivar.     

Morphological studies on the Sulawesi macaques I: Phyletic analysis of body color

The body color of Sulawesi macaques was measured quantitatively and compared among the different monkeys. As a result, divergence models for extant Sulawesi macaques, withtonkeana as the starting point and fading as the sole direction of color change, were inferred as follows: (1) fading slightly on the upper half of the body—nigra, fading more on the proximal part of the body—nigrescens; (2) fading over the whole body—maura; (3) fading greatly on the legs—hecki; and (4) fading on the distal part of the body—ochreata, fading more over the whole body, including the proximal part of the body—brunnescens. The color changed progressively in the order of (1) through (4). The divergence model, excluding the position ofhecki (3), supports the speciation model ofFooden (1969). If the proto-Sulawesi macaques had a body color pattern similar to the livingnemestrina, darkening would have been necessary for the evolution of the Sulawesi macaques after their immigration, and it may have been acquired as an adaptation to the ground (forest floor) living nature of the Sulawesi macaques, together with influences deriving from the insularity and/or from the absence of predators.     

DNA Sequencing Using Ultra Small Amounts of Reagents and Template

One of the key points in the genome project is finding waysto reduce the running cost in DNA sequencing. One way is touse a highly-sensitive fluorescent DNA sequencer, where onlytrace amounts of template DNA and reagents are needed. An experimentalprotocol optimized for the trace amounts of DNA analysis wasestablished by using the hybridization reaction rate coefficientof primers on template DNA, which was estimated to be 7.5x10⁵M^–1 sec^–1 at 37°C. One femtomole of templateDNA with 0.001 unit of modified T7 DNA polymerase (SequenaseVer. 2.0) and also 0.45 fmol of M13 template DNA with 0.01 unitof Taq DNA polymerase were enough to sequence DNA of up to 400bases.     

Endogenous inhibitors of factor B

proteins which were able to bind noncovalently with mouse factor B were found in cells that are nonsecretors of factor B such as mouse-established monocytic cells and L cells but not in peritoneal resident macrophages. These proteins were isolated from lysates of L cells and separated into four distinct proteins by preparative SDS-polyacrylamide gel electrophoresis, with molecular weights of 25K , 28K , 33K, and 35K . The individual proteins formed a complex with purified mouse factor B at a molecular ratio of 1: 1 and inhibited its hemolytic activity. Proteins 25K and 28K inhibited the hemolytic activity of an activated form of factor B combined with cobra venom factor as well as that of the native form. These inhibitors did not affect the hemolytic activity of the second component of complement in mouse serum. The inhibitory activity of the 25K protein was partially inhibited by antiserum raised against it in rabbits.     

The anatomy and clinical applications of the buccal fat pad

The buccal fat pad is an anatomically complex structure that has great importance in facial contour. In properly selected individuals, judicious harvesting of buccal fat can produce dramatic changes in facial appearance by reducing the fullness of the cheek and highlighting the malar eminences. Using fresh cadaver dissection, the anatomy of the buccal fat pad is delineated and its relationship to the masticatory space, facial nerve, and parotid duct is defined. An intraoral approach for buccal fat harvesting is described based on these anatomic findings. Clinical experience manipulating the buccal fat pad for aesthetic modification of facial contour is illustrated.     

Biostereometrics and computergraphics for patients with craniofacial malformations: diagnosis and treatment planning

Utilizing the techniques of biostereometric photography and current state-of-the-art computergraphics, topographic contourgrams have been generated in a series of patients with complex craniofacial malformations. These contourgrams are considered useful in diagnosis and treatment planning and as an aid in postoperative follow-up. We hope that the technique will provide a means to study longitudinal growth in unoperated and operated patients with craniofacial malformations. Comparisons between these groups and normal patients are currently being investigated.