Direct Injection of CRISPR/Cas9-Related mRNA into Cytoplasm of Parthenogenetically Activated Porcine Oocytes Causes Frequent Mosaicism for Indel Mutations

Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the α-1,3-galactosyltransferase that synthesizes the α-Gal epitope using parthenogenetically activated porcine oocytes. The lack of α-Gal epitope expression can be monitored by staining with fluorescently labeled isolectin BS-I-B₄ (IB4), which binds specifically to the α-Gal epitope. When oocytes were injected with guide RNA specific to GGTA1 together with enhanced green fluorescent protein (EGFP) and human Cas9 mRNAs, 65% (24/37) of the developing blastocysts exhibited green fluorescence, although almost all (96%, 23/24) showed a mosaic fluorescent pattern. Staining with IB4 revealed that the green fluorescent area often had a reduced binding activity to IB4. Of the 16 samples tested, six (five fluorescent and one non-fluorescent blastocysts) had indel mutations, suggesting a correlation between EGFP expression and mutation induction. Furthermore, it is suggested that zygote microinjection of mRNAs might lead to the production of piglets with cells harboring various mutation types.     

Development of a hyper-interfacial bonding process for ultrafine-grained steels

With the rapid progress of technology in a wide variety of industries, such as aerospace, IT, civil engineering/construction, energy, etc., the metals used in these fields have recently seen increasingly stringent requirements being imposed on their strength, toughness, corrosion resistance, durability, etc. A capability for resource- and energy-saving as well as good recyclability are also being increasingly sought to counter a number of progressively more serious problems facing the global environment. To meet these challenges, efforts are being mounted to develop so-called super-steels providing a good combination of high strength and high toughness through submicron order refinement of their grain size while retaining a basic mild steel chemistry in order to lessen their environmental impact and reduce total life cycle costs.     

"Pored-domes" of the fenestrated endotheliocyte of the glomerular and peritubular capillaries in the rodent kidney.

The fenestrated endotheliocyte of peritubular and glomerular capillaries in rat and mouse kidneys were observed with SEM and TEM. In the glomerular capillary, so-called "pored-domes" were found not only at the fenestrated areolae but also at the nuclear region of the endotheliocyte. At the region between filtration surface and nuclear region, they accumulated to construct a sponge-like structure. The endotheliocyte of peritubular capillary also showed small "pored-domes". The size and morphology of the pores in the "pored-domes" of glomerular and peritubular capillaries were similar to those of areolae fenestratae of the respective capillary. Based on the findings, we assumed that pored-domes and the sponge-like structure are the reservoir for the fenestrated area of the endotheliocyte to accommodate the rapid expansion of capillary lumen.     

A Study about the Effect of Space Size on the Individual Distance

This study is the design guidelines for the living space with robots. The study can be applied to the architectural planning and focused on the basic environmental primary factor. The authors focused on space size, specifical distance from the wall in the front direction, position, and distance between an individual as factors affecting individual distance. The authors research the distances a small mobile robot moves to a six mats space compared with that toward a distances 5 m or more （space less physical factors）. The following three results were obtained： （1） The individual distance is smaller than 30 cm in both the approach angle in the experiment space of six mats; （2） The distance from the wall in the front increased, and the individual distance between a participant and a robot also increased; （3） In terms of body position, the upright positioning was clarified and the individual distance was small.     

Evaluation of MC3T3-E1 Cell Osteogenesis in Different Cell Culture Media

Many biomaterials have been evaluated using cultured cells. In particular, osteoblast-like cells are often used to evaluate the osteocompatibility, hard-tissue-regeneration, osteoconductive, and osteoinductive characteristics of biomaterials. However, the evaluation of biomaterial osteogenesis-inducing capacity using osteoblast-like cells is not standardized; instead, it is performed under laboratory-specific culture conditions with different culture media. However, the effect of different media conditions on bone formation has not been investigated. Here, we aimed to evaluate the osteogenesis of MC3T3-E1 cells, one of the most commonly used osteoblast-like cell lines for osteogenesis evaluation, and assayed cell proliferation, alkaline phosphatase activity, expression of osteoblast markers, and calcification under varying culture media conditions. Furthermore, the various media conditions were tested in uncoated plates and plates coated with collagen type I and poly-L-lysine, highly biocompatible molecules commonly used as pseudobiomaterials. We found that the type of base medium, the presence or absence of vitamin C, and the freshness of the medium may affect biomaterial regeneration. We posit that an in vitro model that recapitulates in vivo bone formation should be established before evaluating biomaterials.     

Effects of Docosahexaenoic Acid Content in Triacylglycerol on Human Taste Perception

ABSTRACT: To elucidate the contribution of docosahexaenoic acid (DHA) in triacylglycerol to tuna oil specific taste effects, suppression of bitterness and enhancement of umami, we conducted sensory analysis employing an oil-in-water emulsion prepared with taste substances and the oils differed in the DHA content. The tuna oil specific taste effects were linearly dependent on DHA content. The increase of DHA in oil also linearly enhanced umami and flavor (continuity and richness) of synthetic tuna extract. These results indicate that DHA, a major fatty acid of tuna oil, greatly contributes to tuna oil specific effects on basic tastes and flavor.     

Uncertainty of calibration of 2D planar Coordinate Measuring Machine

When the kinematic parameters of 2D planar Coordinate Measuring Machine are calibrated in self-calibration method, it is shown how to estimate the uncertainty of calibration and how to estimate the uncertainty of measurement. In this paper, the uncertainty of probing and the uncertainty of sensors are considered as the contributors of uncertainty. Two different types of CMMs are applied to two different artefacts for calibration and those measures in some different orientations for estimation of uncertainty. The effectiveness of the difference of CMMs, the artefacts and the measurement orientations are compared. As a result, it is proved that the uncertainty distribution is different in the orientation of measurement and the uncertainty could be smaller by selecting measurement orientation.     

Evaluation of the efficacy of orange roughy (Hoplostetbus atlanticus) oil in subjects with dry skin

Moisturizers have beneficial effects in treating dry skin. The objective of this study was to evaluate the effect of orange roughy (Hoplostethus atlanticus) oil, a marine-derived wax ester, on skin dryness in comparison with a reference commercially available petrolatum-based moisturizer (Vaseline) and untreated control. Subjects (n = 24) with moderate to severe skin dryness at the lower limb of legs (Study 1) and with certain degree of skin dryness on the face and the forearms (n = 22, Study 2) were treated twice a day for 42 consecutive days with the test products in randomized clinical trials. Transepidermal water loss (TEWL) was measured at the beginning and the end of Study 1, whereas skin hydration was measured at the beginning, after 3 and 6 weeks of the application (the end of the study) in Study 2. Changes in the skin dryness were assessed by a dermatologist using a video microscopy. In Study 1, the dryness score of skin applied with orange roughy oil improved significantly (P < 0.01) in 6 weeks. The skin looked smooth with no or little dry scaly skin. Orange roughy oil was evaluated with a 60% efficacy in treating skin dryness by the expert, which comparable to that of petrolatum (68%). No significant change in TEWL was found either in orange roughy oil or petrolatum treatment, although the values showed a tendency to improve in both cases. Similarly, the results of the skin capacitance in Study 2 showed a significant improvement of the skin symptoms after 3 and 6 weeks. These results showed that the performance of orange roughy oil in treating skin dryness was comparable to that of petrolatum.