Weproduced a transgenic rodent malaria parasite (Plasmodium berghei) that contained the luciferase gene under apromoter region of elongation factor-1α. These transgenic (TG) parasites expressed luciferase inall stages of their life cycle, as previously reported. However, we were the firstto succeed in observing sporozoites as a mass in mouse skin following theirdeposition by the probing of infective mosquitoes. Our transgenic parasites mayhave emitted stronger bioluminescence than previous TG parasites. The estimatednumbers of injected sporozoites by mosquitoes were between 34 and 775 (median 80). Since luciferase activity diminished immediately after the death of theparasites, luciferase activity could be an indicator of the existence of liveparasites. Our results indicated that sporozoites survived at the probed sitefor more than 42 hours. We also detected sporozoites in the liver within 15 minof the intravenous injection. Apart from the liver, bioluminescence was notobserved in the lung, kidney, or spleen. We reconfirmed that the liver was thefirst organ for malaria parasites to enter and increase in number.

We produced a transgenic rodent malaria parasite (Plasmodium berghei) that contained the luciferase gene under a promoter region of elongation factor-1α. These transgenic (TG) parasites expressed luciferase in all stages of their life cycle, as previously reported. However, we were the first to succeed in observing sporozoites as a mass in mouse skin following their deposition by the probing of infective mosquitoes. Our transgenic parasites may have emitted stronger bioluminescence than previous TG parasites. The estimated number of injected sporozoites by mosquitoes was between 34 and 775 (median 80). Since luciferase activity diminished immediately after the death of the parasites, luciferase activity could be an indicator of the existence of live parasites. Our results indicated that sporozoites survived at the probed site for more than 42 hours. We also detected sporozoites in the liver within 15 min of the intravenous injection. Bioluminescence was not observed in the lung, kidney or spleen. We confirmed the observation that the liver was the first organ in which malaria parasites entered and increased in number.

We assessed the effectiveness of practical instruction in parasitology for undergraduates at Jichi Medical School by examining grades on a practical examination. Two hundred six second-year medical students in 1997 and 1998 (103 students in each year) were enrolled in this study. The students took written and practical examinations at the end of the program. We found that grades on the practical examination were correlated with grades on the written examination (r=0.5664; p<0.001). The discrimination index ranged from 0.23 to 0.78. The percentage of correctly identified species was significantly higher when students studied live specimens than when they used other methods (p<0.0001 in both 1997 and 1998). The highest rates of correct identification (more than 90%) were for Anisakis species larvae and Enterobius vermicularis eggs in 1997 and for Anisakis species larvae, E. vermicularis eggs, and Anopheles mosquitoes in 1998. Results of neither written nor practical examinations differed significantly between students who chose biology at the entrance examination and those who did not. Our results suggest that undergraduates would gain a better understanding of parasitology by studying live specimens.

It has been proposed that transgenic mosquitoes can be used as a “flying syringe” for infectious disease control. We succeeded in generating a transgenic (TG) mosquito, Anopheles stephensi, excreting and discharging DsRed in saliva. DsRed was deposited on the membrane where the TG mosquito probed with its proboscis. Repeated feeding by the TG mosquitoes induced anti-DeRed as well as anti-SG antibodies in mice. This indicates that the TG mosquitoes can immunize the animal. Moreover, in this report, we employed a pre-immunization method before exposing mice to the TG mosquitoes. We injected DsRed to mice to prepare memory B cells and exposed the mice to bites by the TG mosquitoes excreting DsRed. The mice produced a higher titer of antibody to DsRed, suggesting that the bites from TG mosquitoes act as a booster and that primary immunization with a vaccine protein and exposure to TG mosquitoes excreting the vaccine protein in the saliva produces a synergistic effect.

Objective: To convey HIV⁄AIDS-related knowledge to people in rural Cambodia, we conducted an HIV⁄AIDS awareness intervention program and investigated its effectiveness, participants’ sexual behavior, HIV-related knowledge, and their attitude to HIV⁄AIDS.
Methods: We conducted HIV⁄AIDS awareness intervention in a rural area of Cambodia from April to November 2007. We selected three villages (a total of 180 villagers) in Siem Reap Province. Our HIV⁄AIDS awareness intervention involved practical explanations by well-trained Cambodian staff using visual material and participatory activities in order to promote interest among illiterate participants. We implemented a cross-sectional study in each village after the HIV⁄AIDS awareness intervention using a questionnaire written in Khmer and assisted by a Cambodian NGO.
Results: Two-thirds of the participants had not finished primary school and had difficulties reading and writing. A total of 77.8% of the people had obtained HIV⁄AIDS-related information from NGOs.
Conclusion: The HIV⁄AIDS awareness intervention was welcomed by most of the villagers and positively influenced HIV⁄AIDS-related knowledge through the use of practical explanations. Rural areas are still more vulnerable to HIV⁄AIDS transmission, and at the same time more likely to be influenced by NGOs, than cities because of high rates of illiteracy and a lack of access to general HIV⁄AIDS-related information sources including television, books, newspapers, and the Internet. NGOs need to increase their efforts to educate the vulnerable populations in rural areas.

In this study, we examined the feasibility of using subzonal cell injection with electrofusion for interspecies somatic cell nuclear transfer (iSCNT) to produce sei whale embryos and to improve their developmental capacity by investigating the effect of osmolarity and macromolecules in the culture medium on the in vitro developmental capacity. Hybrid embryos produced by the electrofusion of fetal whale fibroblasts with enucleated porcine oocytes were cultured in modified porcine zygote medium-3 to examine the effects of osmolarity and fetal serum on their in vitro developmental capacity. More than 66% of the whale somatic cells successfully fused with the porcine oocytes following electrofusion. A portion (60~81%) of the iSCNT whale embryos developed to the two- to four-cell stages, but no embryos were able to reach the blastocyst stage. This developmental arrest was not overcome by increasing the osmolarity of the medium to 360 mOsm or by the addition of fetal bovine or fetal whale serum. Our results demonstrate that sei whale-porcine hybrid embryos may be produced by SCNT using subzonal injection and electrofusion. The pig oocytes partly supported the remodeling and reprogramming of the sei whale somatic cell nuclei, but they were unable to support the development of iSCNT whale embryos to the blastocyst stage.
Animals ; Cloning, Organism/*veterinary ; Culture Media ; Embryo, Mammalian ; Karyotyping ; Nuclear Transfer Techniques/*veterinary ; *Oocytes ; Swine/*embryology ; Whales/*embryology