To establish better embryo induction procedures in anther culture of apple, the relationships between the microspore stage and the flower bud stage, and between embryo induction and the developmental stage of microspores during cold pre-treatment of flower buds were investigated. We classified the flower bud stages into four: "Tight cluster", "First pink", "Pink", and "Full pink". Each stage was defined by a morphological characteristic. The microspore stages were classified into seven: "tetrads", "early uninucleate", "miduninucleate", "late uninucleate", "early binucleate", "late binucleate", and "mature grain". The morphological characteristic in each stage was clarified. The "Tight cluster" flower bud stage mainly corresponded with the microspore stage of tetrad to mid-uninucleate. The "First pink" of the flower cluster mainly corresponded with the mid-uninucleate and late uninucleate microspore. The "Pink" and "Full pink" of flowering corresponded respectively with the late binucleate and the mature grain. Cold pre-treatment for more than 25 days of flower buds was found to be an efficient means of increasing embryo formation in anther culture. The microspore of late uninucleate and early binucleate stages grew into embryos efficiently after 25 days of cold pre-treatment. To achieve that condition, flower buds at late "Tight cluster" to early "First pink" stage were collected because the microspore stage progresses during 25 days of cold pre-treatment. Most microspores were early uninucleate to early binucleate stages at the "Tight cluster" to "First pink" flower bud stages. The shoots regenerated from embryos were analyzed using SSR markers. The investigation revealed that all shoots had a haploid origin.
We previously developed a new system of magnetic induction interstitial hyperthermia, implant heating system (IHS), and applied to oral cancer control. To overcome the limitations of this system, the present study dealt with new magnetic induction hyperthermia using Magnetic Cationic Liposomes (MCLs) which are injected into a tumor. We prepared a tongue tumor model using the transplantable rabbit VX-7 cells. MCL was injected into the tumor margins to heat the tumor selectively and induce blood vessel damage by heating. Two separated MCL injection sites were made by using Micro syringe pump and sandwiched the tumor on both sides. The rabbit in group I (n = 4) were performed hyperthermia, while those in group II (n= 4) were not subjected to magnetic field irradiation as the control. In the experimental rabbits, the temperature in the tumor was as high as 43.0°C. The tumor volumes of irradiated rabbits were significantly smaller than those of the non irradiated ones. The transplanted tumor of experimental rabbits almost completely disappeared following 18 day after treatment. The tongues, removed from irradiated rabbits on 28th day after irradiation, showed no tumor cells and fibrous tissue replacement completely occurred in the implanted tumor area. Tongue muscle adjacent to the MCL injection sites did not exhibit any evidence of inflammation or necrosis. The present study confirmed the effectiveness of interstitial hyperthermia using MCLs against oral carcinoma.
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