Original Study: Comparison of three intraosseous access techniques in cats

Objective – To compare the placement feasibility and amount of bone trauma induced by 3 intraosseous (IO) access techniques in cats: an automatic impact penetration device (A), an automatic rotary insertion device (B), and a manual IO needle (C). Design – Prospective ex vivo study. Setting – University. Animals – Eighteen adult mixed breed feline cadavers. Interventions – Cadavers provided 72 total IO insertion locations divided equally between the right and left humerus and tibia. The 3 IO techniques were randomly allocated to these locations. Time to successful insertion, ease of insertion, and success rate were recorded. Each insertion site was analyzed for the number of bone fragments and defect diameter by computed tomography. Measurements and Main Results – Device B had lower time of insertion (P=0.01) compared with devices A and C. Device B had better ease of insertion scores (P<0.01) compared with devices A and C. No differences were detected between insertion sites (tibia versus humerus). No differences in the number of bone fragments, defect diameter, or success rate were detected among devices (P=0.06, 0.31, and 0.14, respectively). Conclusions – All 3 IO access methods evaluated yield acceptable results. Device B is significantly faster and easier to place in cat cadavers when compared with other methods.     

High developmental rates of mouse oocytes cryopreserved by an optimized vitrification protocol: the effects of cryoprotectants, calcium and cumulus cells

Unfertilized oocytes are one of the most desired germ cell stages for cryopreservation because these cryopreserved oocytes can be used for assisted reproductive technologies, including in vitro fertilization (IVF) and intracytoplasmic sperm injection. However, in general, the fertility and developmental ability of cryopreserved oocytes are still low. The aim of the present study was to improve vitrification of mouse oocytes. First, the effects of calcium and cryoprotectants, dimethyl sulfoxide and ethylene glycol (EG), in vitrification medium on survival and developmental ability of vitrified oocytes were evaluated. Oocytes were vitrified by a minimal volume cooling procedure using different cryoprotectants. Most of the vitrified oocytes were morphologically normal after warming, but their fertility and development were low independently of calcium and cryoprotectants. Second, the effect of cumulus cells on ability of oocytes to be fertilized and develop in vitro was examined. The fertility and developmental ability of denuded oocytes (DOs) after IVF were reduced compared with cumulus-oocyte complexes (COCs) both in fresh and cryopreserved groups. Vitrified COCs showed significantly (P<0.05) higher fertility and ability to develop to the 2-cell and blastocyst stages than those of vitrified DOs with cumulus cells and vitrified DOs alone. The vitrified COCs developed to term at a high success rate equivalent to the rate obtained with IVF using fresh COCs. Taken together, the current results clearly demonstrate that, in the presence of surrounding cumulus cells, matured mouse oocytes vitrified using calcium-free media and EG retain their developmental competence. These findings will contribute to improve oocyte vitrification in not only experimental animals but also clinical application for human infertility.     

Anticancer activities of thelephantin O and vialinin A isolated from Thelephora aurantiotincta

Thelephora aurantiotincta is an edible mushroom belonging to the genus Thelephora; it grows in symbiosis with pine trees. Recently, phytochemical investigations have revealed that the genus Thelephora is an abundant source of p-terphenyl derivatives. However, their bioactivity has not yet been well characterized. In screening for natural materials with anticancer activity, a T. aurantiotincta ethanol extract (TAE) was found to decrease cell viability in human hepatocellular carcinoma cells (HepG2). In this study, a new p-terphenyl derivative, thelephantin O, and a known compound, vialinin A, were isolated as the principal bioactive components of TAE. These compounds decreased cell viability in HepG2 and human colonic carcinoma cells (Caco2), but not in noncancerous human hepatocytes. This is the first report of the isolation from T. aurantiotincta of selective cytotoxic agents against cancer cells.     

Triterpenoids isolated from Zizyphus jujuba inhibit foam cell formation in macrophages

Because foam cell formation in macrophages is believed to play an essential role in the progression of early atherosclerotic lesions in vivo, prevention of foam cell formation is considered to be one of the major targets for the treatment of atherosclerosis. The present study examined the inhibitory effect of 50 crude plant extracts on foam cell formation. Among those crude extracts, Zizyphi Fructus (ZF) and Zizyphi Semen (ZS) extracts significantly inhibited the foam cell formation induced by acetylated LDL. Furthermore, triterpenoids such as oleanonic acid, pomolic acid, and pomonic acid were the major active compounds, and triterpenoids containing a carboxylic acid at C-28 play an important role in the inhibitory effect on foam cell formation in human macrophages. These data suggest that triterpenoids in Zizyphus jujuba , the plant source of ZF and ZS, may therefore be useful for the prevention of atherosclerosis.     

A Study of Combining Elastin in the Chitosan Electrospinning to Increase the Mechanical Strength and Bioactivity

While electrospun chitosan membranes modified to retain nanofibrous morphology have shown promise for use in guided bone regeneration applications in in vitro and in vivo studies, their mechanical tear strengths are lower than commercial collagen membranes. Elastin, a natural component of the extracellular matrix, is a protein with extensive elastic property. This work examined the incorporation of elastin into electrospun chitosan membranes to improve their mechanical tear strengths and to further mimic the native extracellular composition for guided bone regeneration (GBR) applications. In this work, hydrolyzed elastin (ES12, Elastin Products Company, USA) was added to a chitosan spinning solution from 0 to 4 wt% of chitosan. The chitosan–elastin (CE) membranes were examined for fiber morphology using SEM, hydrophobicity using water contact angle measurements, the mechanical tear strength under simulated surgical tacking, and compositions using Fourier-transform infrared spectroscopy (FTIR) and post-spinning protein extraction. In vitro experiments were conducted to evaluate the degradation in a lysozyme solution based on the mass loss and growth of fibroblastic cells. Chitosan membranes with elastin showed significantly thicker fiber diameters, lower water contact angles, up to 33% faster degradation rates, and up to seven times higher mechanical strengths than the chitosan membrane. The FTIR spectra showed stronger amide peaks at 1535 cm⁻¹ and 1655 cm⁻¹ in membranes with higher concentrated elastin, indicating the incorporation of elastin into electrospun fibers. The bicinchoninic acid (BCA) assay demonstrated an increase in protein concentration in proportion to the amount of elastin added to the CE membranes. In addition, all the CE membranes showed in vitro biocompatibility with the fibroblasts.     

Cloning of the phloem-specific small heat-shock protein from leaves of rice plants

The N-terminal amino-acid sequence of a major rice phloem-sap protein, designated as RPP23, was determined. The complete amino-acid sequence of RPP23 was deduced from the corresponding rice EST- clone and contained an extra 46 amino acids at the N-terminus, that was apparently cleaved off to form mature RPP23 in sieve tubes. RPP23 shared a similarity to plant small heat-shock proteins (smHSPs), though the N-terminal region of RPP23 was distinct from that of known smHSPs. Immunocytological analyses using leaf sections showed that RPP23 was located only in the phloem regions of leaves, and was present in non-stressed plants. In mature leaves, stronger immunocytological signals were detected in sieve elements than in companion cells.     

Microbial mineralization of organic nitrogen into nitrate to allow the use of organic fertilizer in hydroponics

Hydroponics is an excellent technique for the cultivation of vegetable crops and other plants, but organic fertilizers cannot be used in conventional hydroponic systems, which generally use only inorganic fertilizers, because organic compounds in the hydroponic solutions generally have phytotoxic effects that lead to poor plant growth. Few microorganisms are present in hydroponic solutions to mineralize the organic compounds into inorganic nutrients. In this article a novel and practical hydroponic culture method that uses microorganisms to degrade organic fertilizer in the hydroponic solution has been developed. Soil microorganisms were cultured by regulating the amounts of organic fertilizer and inoculum, with moderate aeration. The microorganisms mineralized organic nitrogen via ammonification and nitrification into nitrate at an efficiency of 97.6%. The culture solution containing the microorganisms was usable as a hydroponic solution, and organic fertilizer could be directly added to it during vegetable cultivation. Vegetables grew well in the organic hydroponic system. Organic hydroponics based on this method is therefore a practical tool for the utilization of organic sources of fertilizer.     

Effect of CO2 enrichment on fruit growth and quality in Japanese pear (Pyrus serotina Reheder cv. Kosui)

Six year-old Japanese pear (Pyrus seratina Reheder cv. Kosui) trees grafted on P. serotina cv. Nihonyamanashi were grown in containers filled with Granite Regosol under glasshouse conditions. At different stages of fruit growth, pear trees were exposed to an elevated CO₂ concentration (130 Pa CO₂ ) along with a control (35 Pa CO₂). For one group of plants, CO₂ enrichment was applied for 79 d from 52 d after full bloom (DAB) to fruit maturity (long-term CO₂ enrichment) and for another group the same treatment was applied for 35 d from 96 DAB to fruit maturity (short-term CO₂ enrichment). The effects of the elevated CO₂ concentration on vegetative growth, mineral contents, and fruit production and quality were examined. Long-term CO₂ enrichment enhanced vegetative growth, without any significant effect on the mineral contents in either flower bud or fruit except for a remarkable increase in the K content. Long-term CO₂ enrichment increased the fruit size and fresh weight, but had no significant effect on the fruit quality. On the other hand, the short-term CO₂ enrichment did not induce any significant change in the fruit size but increased the fruit sugar concentration. Along with the reduction of the sorbitol concentration in fruit, the fructose and sucrose concentrations increased and these changes occurred earlier at elevated CO₂ than at ambient CO₂ concentrations. From these results, we concluded that the effect of CO₂ enrichment on fruit growth varies depending upon the growth stages of fruit: during the initial and fruitlet stages when fruit expansion occurs, CO₂ enrichment increases the fruit size, whereas, during maturation when fruit expansion has slowed down and sugar accumulation in fruit is active, it increases the fruit sugar concentration.