Surgical correction of aberrant conjunctival overgrowth in a rabbit: a case report

A dwarf rabbit presented with unilateral aberrant conjunctival growth. Allgoewer’s U-suture therapy was initially used to correct the overgrowth. Centrifugal incisions extending up to the limbus were made on the hypertrophic conjunctiva. Transpalpebral limbal fixation was performed next. When the symptoms recurred 3 weeks later, a second operation was performed using the Lembert suture method instead. The overgrowing membrane was excised radically just posterior to the limbus. The conjunctiva was then sutured using the Lembert pattern. The rabbit recovered with no further complications.     

Neurogenic and cardiomyogenic differentiation of mesenchymal stem cells isolated from minipig bone marrow

The present study investigated the potential of minipig bone marrow-mesenchymal stem cells (BM-MSCs) to differentiate in vitro into neuron- and cardiomyocyte-like cells. Isolated BM-MSCs exhibited a fibroblast-like morphology, expressed CD29, CD44 and CD90, and differentiated into osteocytes, adipocytes and chondrocytes. Upon induction in two different neuronal specific media, most of BM-MSCs acquired the distinctive morphological features and positively stained for nestin, neurofilament-M (NF-M), neuronal nuclei (NeuN), β-tubulin, galactocerebroside (Gal-C) and glial fibrillary acidic protein (GFAP). Expression of nestin, GFAP and NF-M was further demonstrated by RT-PCR and RT-qPCR. Following cardiomyogenic induction, MSCs exhibited a stick-like morphology with extended cytoplasmic processes, and formed cluster-like structures. The expression of cardiac specific markers α-smooth muscle actin, cardiac troponin T, desmin and α-cardiac actin was positive for immunofluorescence staining, and further confirmed by RT-PCR and RT-qPCR. In conclusion, our results showed the in vitro differentiation ability of porcine BM-MSCs into neuron-like and cardiomyocyte-like cells.     

Influence of Seasons, Extenders, Slow and Rapid Freezing on Seminal Characters in Korean Native Bucks

The success of AI technology is based on both semen quality and freezing process. In order to establish the semen freezing techniques in Korean native bucks, factors affecting the success were evaluated in the present study. Semen collected by electro-ejaculation from bucks during four distinct seasons was evaluated for semen volume and pH, sperm motility and survivability. The semen volume, concentration and total cell were higher in spring, summer and less in winter. Yet, there were no seasonal differences in the proportional data of live sperm, motility score and pH of semen among seasons. The percentage of live sperm after thawing was found to increase with increased concentration of lactose in Tris-Egg yolk-glycerol (TY-G), being highest in TY-G supplemented with 180 mm lactose (TYL180-G), but did not differ between TY-G and TYL120-G. Sperm motility was enhanced by employing 2.0 h equilibration time with rapid freezing method. In conclusion, semen could be frozen with high success rates for further use of AI in breeding techniques and to preserve the Korean native bucks.     

Polyacetylenic compounds, ACAT inhibitors from the roots of Panax ginseng

Acyl-CoA: cholesterol acyltransferase (ACAT), which plays a role in the absorption, storage, and production of cholesterol, has been explored as a potential target for pharmacological intervention of hyperlipidemia and atherosclerotic disease. In our search for ACAT inhibitors from natural sources, the petroleum ether extract of Panax ginseng showed moderate inhibition of ACAT enzyme from rat liver microsomes. Bioactivity-guided fractionations led to the isolation of one new polyacetylenic compound, (9R,10S)-epoxy-16-heptadecene-4, 6-diyne-3-one (1), in addition to the previously reported polyacetylenic compounds 2 and 3. Their chemical structures were elucidated on the basis of spectroscopic evidence (UV, IR, NMR, and MS). The compounds 1, 2, and 3 showed significant ACAT inhibition with IC(50) values of 35, 47, and 21 microM, respectively.     

Characterization of Transgenic Pigs That Express Human Decay Accelerating Factor and Cell Membrane‐tethered Human Tissue Factor Pathway Inhibitor

The expression of human complement regulatory proteins (hCRP; hDAF, hCD59, and hMCP) in pig tissues has been suggested as one of strategies to overcome the hyperacute rejection (HAR) in pig‐to‐human transplantation. Expression of human tissue factor pathway inhibitor (hTFPI) in porcine endothelial cells has been suggested as a remedy to overcome microvascular thrombosis. To investigate the effects of these combined transgenes, we established transformed pig cells expressing human decay accelerating factor (hDAF) under the control of enhancer promoter (5′LTR‐PCMVIE), and the fusion protein (hTFPI/hCD4) consisting of the functional domains (K1 and K2) of hTFPI and membrane‐tethering domains (D3 and D4) of hCD4 under the control of PCMVIE. Transgenic pigs were generated with the transformed porcine cells through somatic cell nuclear transfer (SCNT) technology. Analysis of quantitative PCR and real‐time quantitative RT‐PCR showed that four copies of hDAF were integrated and 391 copies of hDAF mRNA expressed in the cells of the transgenic pig. The enhancing activity of 5′LTR was approximately 2 fold compared to CMVIE promoter only. The cell viability test showed that more than 80% of ear cells were viable in the presence of 50% human serum. The chromogenic substrate assay and immunocytochemical staining with tail cells showed that the TFPI activity of fusion protein was observed on the cell membrane. The membrane localization of hDAF and hTFPI proteins was observed by immunocytochemical staining, and the expression of transgenes in heart and liver tissues was also confirmed by immunohistochemistry.     

Comparison of bare metal stent and paclitaxel-eluting stent using a novel rat aorta stent model

The purpose of our study was to create a novel rat aorta stent implantation model. Stainless steel bare metal stents (BMS) or paclitaxel-eluting stents (PES) were implanted in male Sprague-Dawley rats (BW 400 ± 20 g). Two and four weeks after stent implantation, the aorta were collected, fixed with 2% glutaraldehyde, and cut into two segments. One segment was used for scanning electron microscopy analysis to evaluate re-endothelialization, and the other segment was used to calculate the neointimal area. At 2 weeks after stenting, the appearance of neointimal hyperplasia was less in the PES group than in the BMS group. At 4 weeks after stenting, no significant difference in neointimal hyperplasia was observed between two groups. On the other hand, the PES group showed more thrombus formation and less re-endothelialization compared to the BMS group. This study demonstrated the ability of a novel rat model of aorta stenting via a common carotid artery to measure the efficacy and safety of commercially available drug-eluting stents.     

Combined effects of leaf/fruit ratios and N and K fertigation levels on growth and distribution of nutrients in pot-grown persimmon trees

Responses of pot-grown persimmon trees (Diospyros kaki cv. ‘Fuyu’) with two different leaf/fruit (L/F) ratios and three different fertigation levels were studied. On July 2, the L/F ratios of 4-year-old trees were adjusted to 20 and 10 by fruit thinning. From July 4 to August 26, each pot was fertigated with a solution containing 0–0 g (none), 10–8 g (medium), or 20–16 g (high) of nitrogen (N) and potassium (K), respectively. The L/F ratio of 10 resulted in higher yields but smaller fruits containing lower soluble solids than the ratio of 20. The non-fertigated trees with the L/F ratio of 10 produced the smallest fruits, but the medium fertigation increased the size of the fruit at this ratio. The high fertigation level decreased fruit skin color at both ratios. Fruit dry weight increased the most among the tree components, accounting for 48–57% of the tree total dry weight at the L/F ratio of 20 and for 64–72% at the ratio of 10. The increase in tree total dry weight at the ratio of 10 was greater than that at the ratio of 20. The trees receiving medium fertigation had higher tree total dry weights than those receiving high fertigation, especially at the ratio of 10. As the L/F ratio changed to 10 from 20, the percentages of N and K partitioned to fruits in the non-fertigated trees increased from 38% to 51% and from 67% to 96%, respectively, of the tree total N and K increases; the percentage decreased with increasing fertigation level. Although fertigation significantly increased total N and K contents of the tree, the increase in K at the ratio of 10 was less in high fertigation rather than in the medium level. The results indicated that adjusting supplemental N and K was necessary to ensure fruit growth and N and K accumulation for trees with high fruit loads, but high levels of fertigation are not necessarily preferable.