Therapeutic angiogenesis by transplantation of induced pluripotent stem cell-derived Flk-1 positive cells

<p>Abstract</p> <p>Background</p> <p>Induced pluripotent stem (iPS) cells are the novel stem cell population induced from somatic cells. It is anticipated that iPS will be used in the expanding field of regenerative medicine. Here, we investigated whether implantation of fetal liver kinase-1 positive (Flk-1⁺) cells derived from iPS cells could improve angiogenesis in a mouse hind limb model of ischemia.</p> <p>Results</p> <p>Flk-1⁺cells were induced from iPS cells after four to five days of culture. Hind limb ischemia was surgically induced and sorted Flk-1⁺cells were directly injected into ischemic hind limbs of athymic nude mice. Revascularization of the ischemic hind limb was accelerated in mice that were transplanted with Flk-1⁺cells compared with control mice, which were transplanted with vehicle, as evaluated by laser Doppler blood flowmetry. Transplantation of Flk-1⁺cells also increased expression of VEGF mRNA in ischemic tissue compared to controls.</p> <p>Conclusions</p> <p>Direct local implantation of iPS cell-derived Flk-1⁺cells would salvage tissues from ischemia. These data indicate that iPS cells could be valuable in the therapeutic induction of angiogenesis.</p

Regulation of Epithelial Sodium Transport via Epithelial Na+ Channel

Renal epithelial Na+ transport plays an important role in homeostasis of our body fluid content and blood pressure. Further, the Na+ transport in alveolar epithelial cells essentially controls the amount of alveolar fluid that should be kept at an appropriate level for normal gas exchange. The epithelial Na+ transport is generally mediated through two steps: (1) the entry step of Na+ via epithelial Na+ channel (ENaC) at the apical membrane and (2) the extrusion step of Na+ via the Na+, K+-ATPase at the basolateral membrane. In general, the Na+ entry via ENaC is the rate-limiting step. Therefore, the regulation of ENaC plays an essential role in control of blood pressure and normal gas exchange. In this paper, we discuss two major factors in ENaC regulation: (1) activity of individual ENaC and (2) number of ENaC located at the apical membrane

A Preclinical Evaluation towards the Clinical Application of Oxygen Consumption Measurement by CERMs by a Mouse Chimera Model.

We have developed an automated device for the measurement of oxygen consumption rate (OCR) called Chip-sensing Embryo Respiratory Measurement system (CERMs). To verify the safety and the significance of the OCR measurement by CERMs, we conducted comprehensive tests using a mouse model prior to clinical trials in a human in vitro fertilization (IVF) program. Embryo transfer revealed that the OCR measured by CERMs did not compromise the full-term development of mice or their future fertility, and was positively correlated with adenosine triphosphate (ATP) production and the mitochondrial membrane potential (&Delta;&Psi;m), thereby indirectly reflecting mitochondrial oxidative phosphorylation (OXPHOS) activity. We demonstrated that the OCR is independent of embryo morphology (the size) and number of mitochondria (mitochondrial DNA copy number). The OCR correlated with the total cell numbers, whereas the inner cell mass (ICM) cell numbers and the fetal developmental rate were not. Thus, the OCR may serve as an indicator of the numbers of trophectoderm (TE) cells, rather than number or quality of ICM cells. However, implantation ability was neither correlated with the OCR, nor the embryo size in this model. This can probably be attributed to the limitation that chimeric embryos contain non-physiological high TE cells counts that are beneficial for implantation. CERMs can be safely employed in clinical IVF owing to it being a safe, highly effective, non-invasive, accurate, and quantitative tool for OCR measurement. Utilization of CERMs for clinical testing of human embryos would provide further insights into the nature of oxidative metabolism and embryonic viability

専門看護師の看護実践能力向上に向けた聖隷CNS 事例検討会の活動について

紀要委員会企画Special Articles　専門看護師（CNS：Certified Nurse Specialist、以下CNS とする） 制度は日本看護協会が1994 年から開始した制度である。本学にはがん看護学、老年看護学、慢性看護学、急性看護学、小児看護学領域にCNS 教育課程が設けられ、さらに2019 年度には在宅看護学領域にも開設を予定している。本学では大学院修了生に対する支援体制を整備するために、2010 年からがん看護学領域の修了生を中心として「聖隷がん看護事例検討会」を立ち上げ、現在は他領域の修了生やCNS を含めた「聖隷CNS 事例検討会」として活動している。本稿では、事例検討会の活動概要、参加者による事例検討会の評価について報告する。今後の事例検討会の発展に向けて、看護学研究科教員への参加依頼、修了生や在学生への働きかけ、事例提供者および参加者の負担軽減策の検討、修了生と教員との連携強化が必要であると考えられる

Comparative Angiogenic Activities of Induced Pluripotent Stem Cells Derived from Young and Old Mice

Advanced age is associated with decreased stem cell activity. However, the effect of aging on the differentiation capacity of induced pluripotent stem (iPS) cells into cardiovascular cells has not been fully clarified. We investigated whether iPS cells derived from young and old mice are equally capable of differentiating into vascular progenitor cells, and whether these cells regulate vascular responses in vivo. iPS cells from mouse embryonic fibroblasts (young) or 21 month-old mouse bone marrow (old) were used. Fetal liver kinase-1 positive (Flk-1+) cells, as a vascular progenitor marker, were induced after 3 to 4 days of culture from iPS cells derived from young and old mice. These Flk-1+ cells were sorted and shown to differentiate into VE-cadherin+ endothelial cells and α-SMA+ smooth muscle cells. Tube-like formation was also successfully induced in both young and old murine Flk-1+ cells. Next, hindlimb ischemia was surgically induced, and purified Flk-1+ cells were directly injected into ischemic hindlimbs of nude mice. Revascularization of the ischemic hindlimb was significantly accelerated in mice transplanted with Flk-1+ cells derived from iPS cells from either young or old mice, as compared to control mice as evaluated by laser Doppler blood flowmetry. The degree of revascularization was similar in the two groups of ischemic mice injected with iPS cell-derived Flk-1+ cells from young or old mice. Transplantation of Flk-1+ cells from both young and old murine iPS cells also increased the expression of VEGF, HGF and IGF mRNA in ischemic tissue as compared to controls. iPS cell-derived Flk-1+ cells differentiated into vascular progenitor cells, and regulated angiogenic vascular responses both in vitro and in vivo. These properties of iPS cells derived from old mice are essentially the same as those of iPS cells from young mice, suggesting the functionality of generated iPS cells themselves to be unaffected by aging

In vitro potency, in vitro and in vivo efficacy of liposomal alendronate in combination with γδ T cell immunotherapy in mice

Nitrogen-containing bisphosphonate (N-BP), including zoledronic acid (ZOL) and alendronate (ALD), have been proposed as sensitisers in γδ T cell immunotherapy in pre-clinical and clinical studies. Therapeutic efficacy of N-BPs is hampered by their rapid renal excretion and high affinity for bone. Liposomal formulations of N-BP have been proposed to improve accumulation in solid tumours. Liposomal alendronate (L-ALD) has been suggested as a suitable alternative to liposomal ZOL (L-ZOL), due to unexpected mice death experienced in pre-clinical studies with the latter. Only one study so far has proven the therapeutic efficacy of L-ALD, in combination with γδ T cell immunotherapy, after intraperitoneal administration of γδ T cell resulting in delayed growth of ovarian cancer in mice. This study aims to assess the in vitro efficacy of L-ALD, in combination with γδ T cell immunotherapy, in a range of cancerous cell lines, using L-ZOL as a comparator. The therapeutic efficacy was tested in a pseudo-metastatic lung mouse model, following intravenous injection of γδ T cell, L-ALD or the combination. In vivo biocompatibility and organ biodistribution studies of L-BPs were undertaken simultaneously. Higher concentrations of L-ALD (40–60 μM) than L-ZOL (3–10 μM) were required to produce a comparative reduction in cell viability in vitro, when used in combination with γδ T cells. Significant inhibition of tumour growth was observed after treatment with both L-ALD and γδ T cells in pseudo-metastatic lung melanoma tumour-bearing mice after tail vein injection of both treatments, suggesting that therapeutically relevant concentrations of L-ALD and γδ T cell could be achieved in the tumour sites, resulting in significant delay in tumour growth

Generation of Indian ink painting image from two-dimensional image data

Recently, non-photorealistic image rendering (NPR) has become the subject of research and development by many researchers due to a rapid improvement in computer graphics. NPR has been used for communication by emphasizing an image or to simulate existing painting techniques. In this paper, we propose a generation method for Indian ink painting image which is an Oriental painting technique from two-dimensional realistic image. In general, Indian ink painting has three kinds of drawing techniques, “Senbyo “ technique to draw only contours of objects, “Mokkotu“ technique to draw only the interior of objects and “Senzen” technique to draw both contours and the interior of objects. In this paper, we simulate Indian ink painting by “Senzen” technique. We extract the contours from two-dimensional realistic image and decide the interior of the object in order to generate the image which is drawn by “Senzen” technique. The final image is generated by combining the inner area of the object with the contours. We generated several Indian ink painting images by our method. The experimental results show that the proposed method is effective for Indian ink painting image rendering

Substitutional and interstitial impurity p-type doping of thermoelectric Mg2Si: a theoretical study

The narrow-gap magnesium silicide semiconductor Mg2Si is a promising mid-temperature (600–900 K) thermoelectric material. It intrinsically possesses n-type conductivity, and n-type dopants are generally used for improving its thermoelectric performance; however, the synthesis of p-type Mg2Si is relatively difficult. In this work, the hole doping of Mg2Si with various impurity atoms is investigated by performing first principles calculations. It is found that the Ag-doped systems exhibit comparable formation energies ΔE calculated for different impurity sites (Mg, Si, and interstitial 4b ones), which may explain the experimental instability of their p-type conductivity. A similar phenomenon is observed for the systems incorporating alkali metals (Li, Na, and K) since their ΔE values determined for Mg (p-type) and 4b (n-type) sites are very close. Among boron group elements (Ga and B), Ga is found to be favorable for hole doping because it exhibits relatively small ΔE values for Si (p-type) sites. Furthermore, the interstitial insertion of Cl and F atoms into the crystal lattice leads to hole doping because of their high electronegativity

Distinct Action of Flavonoids, Myricetin and Quercetin, on Epithelial Cl − Secretion: Useful Tools as Regulators of Cl − Secretion

Epithelial Cl − secretion plays important roles in water secretion preventing bacterial/viral infection and regulation of body fluid. We previously suggested that quercetin would be a useful compound for maintaining epithelial Cl − secretion at a moderate level irrespective of cAMP-induced stimulation. However, we need a compound that stimulates epithelial Cl − secretion even under cAMP-stimulated conditions, since in some cases epithelial Cl − secretion is not large enough even under cAMP-stimulated conditions. We demonstrated that quercetin and myricetin, flavonoids, stimulated epithelial Cl − secretion under basal conditions in epithelial A6 cells. We used forskolin, which activates adenylyl cyclase increasing cytosolic cAMP concentrations, to study the effects of quercetin and myricetin on cAMP-stimulated epithelial Cl − secretion. In the presence of forskolin, quercetin diminished epithelial Cl − secretion to a level similar to that with quercetin alone without forskolin. Conversely, myricetin further stimulated epithelial Cl − secretion even under forskolin-stimulated conditions. This suggests that the action of myricetin is via a cAMPindependent pathway. Therefore, myricetin may be a potentially useful compound to increase epithelial Cl − secretion under cAMPstimulated conditions. In conclusion, myricetin would be a useful compound for prevention from bacterial/viral infection even under conditions that the amount of water secretion driven by cAMP-stimulated epithelial Cl − secretion is insufficient