No abstract

Mesenchymal stem cells (MSCs) promote therapeutic angiogenesis to cure serious vascular disorders. However, their survival period and cytokine-secretory capacity are limited. Although hepatocyte growth factor (HGF) can accelerate the rate of angiogenesis, recombinant HGF is limited because of its very short half-life (<3–5 minutes). Thus, continuous treatment with HGF is required to obtain an effective therapeutic response. To overcome these limitations, we produced genome-edited MSCs that secreted HGF upon drug-specific induction. The inducible HGF expression cassette was integrated into a safe harbor site in an MSC chromosome using the TALEN system, resulting in the production of TetOn-HGF/human umbilical cord blood-derived (hUCB)-MSCs. Functional assessment of the TetOn-HGF/hUCB-MSCs showed that they had enhanced mobility upon the induction of HGF expression. Moreover, long-term exposure by doxycycline (Dox)-treated TetOn-HGF/hUCB-MSCs enhanced the anti-apoptotic responses of genome-edited MSCs subjected to oxidative stress and improved the tube-formation ability. Furthermore, TetOn-HGF/hUCB-MSCs encapsulated by arginine-glycine-aspartic acid (RGD)-alginate microgel induced to express HGF improved in vivo angiogenesis in a mouse hindlimb ischemia model. This study showed that the inducible HGF-expressing hUCB-MSCs are competent to continuously express and secrete HGF in a controlled manner. Thus, the MSCs that express HGF in an inducible manner are a useful therapeutic modality for the treatment of vascular diseases requiring angiogenesis.

Exosomes are nanosized vesicles commonly found in biological fluids as a result of a secretion process involving endosomes and multivesicular bodies. The isolation and analysis of exosomes can be useful for noninvasive clinical diagnosis of a variety of human diseases. We investigated the utility of analyzing exosomal proteins, using matrix-assisted laser desorption/ionization combined with Fourier-transform ion cyclotron resonance mass spectrometry (MALDI-FTICR-MS), as a means of determining the presence of exosomes. MALDI-FTICR-MS analyses of exosomes enriched from human serum via centrifugation in a mass range of m/z 1000–20 000 yielded a distinctive protein around m/z 7766. The high mass accuracy and resolution of MALDI-FTICR-MS allowed for reliable comparisons against a protein database, through which the protein was identified as platelet factor 4 (PLF4), whose singly charged protein peak has an elemental composition of C341H577N96O101S4 +, with a theoretical most abundant isotopic peak at m/z 7765.194 and a theoretical average peak at m/z 7766. The MALDI-TOF MS analysis of exosomes from the serum of 27 patients with different states of liver diseases provided the most abundant PLF4 peak for each mass spectrum, along with several additional minor peaks. In conclusion, MALDI-MS is suitable as an alternative exosome detection method, serving as a valuable confirmation tool, greatly decreasing the time and workload associated with exosome identification.

Abstract. Small phytoplankton are anticipated to be more important in a recently warming and freshening ocean condition. However, little information on the contribution of small phytoplankton to overall phytoplankton production is currently available in the Amundsen Sea. To determine the contributions of small phytoplankton to total biomass and primary production, carbon and nitrogen uptake rates of total and small phytoplankton were obtained from 12 productivity stations in the Amundsen Sea. The daily carbon uptake rates of total phytoplankton averaged in this study were 0.42 g C m −2 d −1 (SD = ± 0.30 g C m −2 d −1 ) and 0.84 g C m −2 d −1 (SD = ± 0.18 g C m −2 d −1 ) for nonpolynya and polynya regions, respectively, whereas the daily total nitrogen (nitrate and ammonium) uptake rates were 0.12 g N m −2 d −1 (SD = ± 0.09 g N m −2 d −1 ) and 0.21 g N m −2 d −1 (SD = ± 0.11 g N m −2 d −1 ), respectively, for non-polynya and polynya regions, all of which were within the ranges reported previously. Small phytoplankton contributed 26.9 and 27.7 % to the total carbon and nitrogen uptake rates of phytoplankton in this study, respectively, which were relatively higher than the chlorophyll a contribution (19.4 %) of small phytoplankton. For a comparison of different regions, the contributions for chlorophyll a concentration and primary production of small phytoplankton averaged from all the non-polynya stations were 42.4 and 50.8 %, which were significantly higher than those (7.9 and 14.9 %, respectively) in the polynya region. A strong negative correlation (r 2 = 0.790, p<0.05) was found between the contributions of small phytoplankton and the total daily primary production of phytoplankton in this study. This finding implies that daily primary production decreases as small phytoplankton contribution increases, which is mainly due to the lower carbon uptake rate of small phytoplankton than large phytoplankton.

Abstract. The in situ primary production rates and various environmental variables were investigated in the Chukchi Sea during the RUSALCA expedition, which was conducted in 2012, to identify the current status of primary production. A 13 C-15 N dual-tracer technique was used to measure the daily primary production rates, which ranged from 0.02 to 1.61 g C m −2 d −1 (mean ±SD = 0.42 ± 0.52 g C m −2 d −1 ). The primary production rates showed large regional differences, with the southern region (0.66 ± 0.62 g C m −2 d −1 ) producing approximately 5 times as much as the northern region (0.14 ± 0.10 g C m −2 d −1 ), which was primarily due to the differences in phytoplankton biomasses induced by regional nutrient conditions. The primary production rates in the Chukchi Sea were averaged using data acquired during the three different RUSALCA expeditions (2004, 2009, and 2012) as 0.33 g C m −2 d −1 (SD = 0.40 g C m −2 d −1 ), which was significantly lower than previously reported rates. In addition to strong seasonal and interannual variations in primary production, recent decreases in the concentrations of major inorganic nutrients and chlorophyll a could be among the reasons for the recent low primary production in the Chukchi Sea because the primary production is mainly affected by nutrient concentration and phytoplankton biomass. The nutrient inventory and primary production appear to be largely influenced by the freshwater content (FWC) variability in the region due to the significant relationships between FWC, nitrate inventory (r = 0.54, p < 0.05), and primary production rates (r = 0.56, p < 0.05). Moreover, we found highly significant relationships between the nutrient inventory and the primary production rates (r = 0.75, p < 0.001). In conclusion, the primary production in the Chukchi Sea is primarily controlled by nutrient availability, which is strongly related to the FWC variability. Our results imply that the predicted increase in freshwater accumulation might cause a decrease in primary production by lowering the nutrient inventory in the euphotic zone of the Chukchi Sea.