In order to determine the susceptible age of Enterobius vermicular is to anthelmintics and to observe the chronologie growth of female E. vermicularis in man, experimental infections were done. About 500 eggs were challenged to 19 volunteers. After 4, 8, 16, 20, 24, 28, 32 and 35 days of infection, each case was treated by either mebendazole or pyrantel pamoate. On the 40th day of infection all cases including control were treated again to terminate the experimental infection and to evaluate the effect of previous treatment. Each case collected 3-day stools to harvest the expelled worms. The results could be summarized as follows: 1. The infection rates of females were in range of 0.6~13.1 % in control cases. Because the collected worms showed comparable growth and development by day, the worms were concluded to be derived from experimental infection. 2. Cases that were treated with mebendazole on 4, 8 and 16 days after infection expelled 37.5%, 2.5% and 67.5% of the number expelled by a control case on the 40th day. Cases treated thereafter expelled no worms on the 40th day. 3. Cases that were treated with pyrantcl pamoates on 4, 8, 16, 24, 28, 32 and 35 days, expelled 90.7%, 25%, 45.3%, 8%, 2.7%, 5% and 29.3% of the number collected from control cases in respect. 4. All the worms collected were females. The total body length increased consistently and comparably from the 20th day of infection. Those collected on the 20th day were 2.5~3.0 mm long with vagina, sac-like structure and strands of ovaries; 24 day-old worms may have short uterus, 28 day-old worms had long uterus without eggs, 32 day-old worms began to produce eggs, 35 day-old worms showed wide variations in egg deposit in uterus, and 40 day-old worms had uterus filled with eggs from vulva to anal levels. From the above results, it was inferred that the life span of female Enterobius vermicularis was longer than 40 days, and the developmental stages of worms younger than 16 days resisted considerably to both mebendazole and pyrantel pamoate.
The effects of cytochalasin B was studied for the cleavage and development of in vitro matured porcine follicular oocytes. The follicular oocytes were collected from slaughtered pig ovaries and matured for 65 hours. The matured oocytes were activated by 7% ethanol(v/v) in DPBS and the activated oocytes were subjected to cytochalasin B concentrations of 2.5, 5.0 and $7.5\;{\mu}g/mL$ for 3, 5 and 7 hours, and then the treated oocytes were cultured in NCSU23 with 0.4% BSA for 7 days. The cleavage rates were not different significantly in each treatment. However, the oocytes treated with $5.0\;{\mu}g/mL$ for 5 hours yielded a significantly higher morula rate(19.7%) than oocytes treated with $2.5\;{\mu}g/mL$ for 3 and 5 hours(9.4%). The sum rate of $2.5\;{\mu}g/mL$ concentration(10.5%) by hour was also significantly lower than those of 5.0(18.0%) and $7.5\;{\mu}g/mL$ concentration(14.6%). The blastocyst rate in oocytes treated with $5.0\;{\mu}g/mL$ for 3(9.4%) and 5 hours(9.0%) was significantly higher than the rate in oocytes treated with $2.5\;{\mu}g/mL$ for 3 hours(0%). The sum rate of $5.0\;{\mu}g/mL$ concentration also significantly higher than those of 2.5 and $7.5\;{\mu}g/mL$ concentration. The results demonstrated that the treatment of oocytes with cytochalasin B of $5.0\;{\mu}g/mL$ for $3{\sim}5$ hours was the optimal concentration and duration for parthenogenetic activation and blastocyst formation of in vitro matured porcine oocytes.
Placenta has been shown to be a site of expression of several of the monoamine membrane uptake transporters. However, the correlation between the expressions of norepinephrine transporter (NET) and placental development including gynecological diseases is still unknown. To investigate the expression and functions of NET in placenta, we conducted to compare NET expression in normal and preeclamptic placenta and analyzed the function of NET in HTR8-SV/neo trophoblast cells after NET gene transfection. The expression of NET was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with preeclamptic placeneta (n=15); and 3) pre-term with preeclamptic placenta (n=11) using semi-quantitative RT-PCR, immunohistochemistry, and Western blot. In order to evaluate the function of NET, NET gene plasmid and NET gene-specific siRNA were trnasfected into HTR-8/SVneo trophoblast cells for 24 hours. NET had low expression in the pre-eclamptic placenta compare with normal placenta but no difference in western blot data. NET was expressed in the trophoblasts, and the up-regulation of NET gene stimulated the invasion of HTR-8/SVneo trophoblast cells by 2.5 fold (p<0.05), whereas the NET-siRNA treatment reduced invasion rates. Also, we observed that the expression of NET induces to expression and activity of MMP-9 in HTR-8/SVneo trophoblast cells in zymography. The results suggest that the expression of NET were reduced in pre-eclampsia and should be inhibited invasion activity of trophoblasts. Therefore, these findings provide useful guidelines for the mechanisms of trophoblast invasion as well as for the basic understanding of gynecological diseases including pre-eclampsia.
The transgenic mice carrying human Interleukin-10 (hIL-10) gene in conjunction with bovine (3 -casein promoter express hIL-10 in milk during lactation. In this study, stability of germ line transmission and expression of hIL-10 transgene integrated into host chromosome were monitored up to generation F8 of transgenic mice. When male mouse of generation F8 was crossbred with normal females, approximately half of offspring (50.9±5.8%) were identified as transgenic mice. Generation F9 to F15 mice also showed similar transmission rates (66.0±20.1%, 61.5±16.7%, 41.1±8.4%, 40.7±20.3%, 61.3±10.8%, 49.2±18.8% and 43.8±25.9%, respectively), implying that hIL-10 transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human IL-10 from milk of generation F9 to F14 mice were 3.6± 1.2 mg/ml, 4.2±0.9 mg/ml, 5.7±1.5 mg/ml, 6.3±3.5 mg/ml, 6.8±4.5 mg/ml and 6.8±3.1 mg/ml, respectively, which was showed high-level expression compared with that of generation F1 (1.6 mg/ml) mice. In conclusion, our results suggest that transgenic mice can be continuously passed their transgenes to the progeny through the breeding program with the same productivity of human IL-10 protein in their milk.
In order to estimate energy budgets of Palaemon macrodactylus, larvae of the shrimp were reared in the laboratory at constant conditions $(25^{\circ}C: 31-32\%o),$ and then juvenile to adult of the shrimp were reared at $15^{\circ}C\;and\;25^{\circ}C$ in the laboratory. Energy used by the reared shrimps were calculated from estimates of data on feeding, growth, molting, metabolism, nitrogen excretion, and energy content. Juveniles and adults reared in the laboratory, which fed on Artemia nauplii, had an average daily growth rates of 0.079 mm/day at $15^{\circ}C\;and\;of\;0.122mm/day\;at\;25^{\circ}C$. The average growth factor* of P. macrodactylus males and females ranged from $3.2\%$ for adult to $13.2\%$ for juveniles individuals, respectively. Intermolt periods were related to body size of the shrimp and to temperature. Average laboratory growth curves were calculated from data on growth factors and intermolt periods to body size of the shrimp at $15^{\circ}C\;and\;25^{\circ}C$. The calorie contents of the shrimp, their molts, eggs and larvae were determined by biochemical composition and oxygen bomb calorimetry. The average amount of energy used in growth for larvae and juvenile to adult were 4.94 cal and 4.55 cal per dry weight in milligram, respectively. The ammount of oxygen used in metabolism was calculated from size, temperature-specific respiration rate. To convert the ammount of oxygen used in respiration into the equivalent energy lost heat was estimated from the data on chemical composition for the larvae and adult, the values was 4.58 cal/ml $O_2$. The energy content per egg was 0.078 cal. The assimilation efficiency estimated by nitrogen content of food and egested faeces gave $61.5\%$ for the larvae. The efficiencies for juvenile to adult ranged between $79.4\%$ and $90.1\%$ The gross growth efficiencies $(K_1)$ and net growth efficiencies $(K_2)$ of P macrodactylus showed $18.33\%\;and 32.63\%$ for total larval stages, ranged from $21.30\%\;to\;31.04\%\;and\;from\;30.03\%\;to\;39.34\%$ for juvenile to adult, respectively.
Purpose of the present study was to find the optimal ovulation induction medicine for the maturation and development of immature oocytes and culture media for 2-cell embryos in the mouse model. ICR female mouse aged 6 to 8 weeks, were stimulated with 5 IU PMSG injection. At 47 to 50 hour post-PMSG injection, ovaries were dissected out and oocytes-cumulus complexes were punctured. The oocyte-cumulus complexes were cultured in media containing various ovulation induction medicine, CC, HMG and Metrodin for 18 hours. Female ICR mice were stimulated with 5 IU PMSG and 48 hours later were injected 5 IU of hCG, then female and male mice were mated. At 48 hour post-hCG injection, oviducts were dissected out and 2-cell embryos were flushed. The 2-cell embryos were cultured in various media, Ham's F-10 media of milli-Q water $(3^{\circ})$, Ham's F-10 media of HPLC (high performance liquid chromatography, Baxter) water, Medicult media, HTF (human tubal fluid) media for 96 hours. The results were as follows. 1. When the oocytes-cumulus complexes were cultured in $10^{-9}{\mu}g/ml{\sim}10^{-8}{\mu}g/ml$ of CC, those were suppressed in meiotic maturation $(28.2{\sim}33.7%)$. Whereas the oocytes-cumulus complexes were cultured in $10^{-7}{\mu}g/ml{\sim}10^{-4}{\mu}g/ml$, these were not effected in meiotic maturation $(54.5{\sim}72.7%)$. 2. When the oocytes-cumulus complexes were cultured in $10^{-4}{\mu}g/ml{\sim}10^{-1}{\mu}g/ml$ of Metrodin, those were suppressed in meiotic maturation $(35.7{\sim}41.5%)$. Meanwhile the oocytes-cumulus complexes were cultured in $10^{-7}{\mu}g/ml{\sim}10^{-5}{\mu}g/ml$, those were not effected in meiotic maturation $(54.2{\sim}70.3%)$. 3. When the oocytes-cumulus complexes were cultured in $10^{-5}{\mu}g/ml{\sim}10^{-4}{\mu}g/ml$ of HMG, those were suppressed in meiotic maturation $(48.2{\sim}50.4%)$. As being cultured in $10^{-7}{\mu}g/ml{\sim}10^{-6}{\mu}g/ml$, increased in meiotic maturation $(75.8{\sim}80.7%)$. 4. When the 2-cell embryos were cultured in Ham's F-10 media of milli-Q water $(3^{\circ})$, Ham's F-10 media of HPLC (high performance liquid chromatograpy, Baxter) water, Medicult media, HTF (human tubal fluid) media, developmental rates to blastocyst and hatching for 96 hour were 50.0%, 45.2%, 71.5% and 95.6%, respectively.
Production of a mature oocyte is a complex process that requires the close association between oocyte and follicular cells. The present study was conducted to investigate the difference between oocytes with and without close junctional communication with cumulus cells and the involvement of two connexins(CXs) in the interactions. Follicles at different sizes(small: 200~400 ㎛; large:>450 ㎛) were mechanically isolated from PMSG-primed mouse ovaries, and punctured to get cumulus-oocyte complex(COC). Oocytes were released themselves(denuded), with partially attached(partial), or with tightly attached(intact) cumulus cells. Maturation and fertilization capacity of the COCs were measured. Expression of CX 37 and CX 43 was examined by RT-PCR and in situ hybridization. The ratio between intact COC and denude/partial oocytes was 30%(SI) and 70%(SPD) in small follicles, while 55%(LI) and 45%(LPD) in large follicles, respectively. Maturation and fertilization rates of the released oocytes were similar among SI, LPD, LI groups, but those were significantly lower in SPD oocytes. In oocytes, CX 37 was the major CX and CX 43 was not expressed, whereas in the cumulus cells, CX 43 was the major, and CX 37 was the next. Results of the present study suggest that 1) immature oocytes from small follicles with intact cell-to-cell communication with cumulus have the similar quality to that of the oocytes from larger follicles, 2) gap junction between oocyte and cumulus cells may be the heterotypic channel, and 3) we could not explain the difference in the cell-to-cell communication between intact and partial/denuded COCs through the expression of the two CXs.
Kim, Seul Gi;Lee, Su Mi;Park, Jun Hong;Song, Kuk;Shin, Byung Sik
Korean Journal of Environmental Biology
/
v.34
no.4
/
pp.346-352
/
2016
To investigate the oviposition preference and development of azuki bean weevil (Callosobruchus chinensis L.), the following six different leguminous seeds were used in this study: red bean (Vigna angularis (Willd.) Ohwi & Ohashi), black soybean (Glycine max (L.) Merr.), soybean (Glycine max (L.) Merr.), seoritae (Glycine max (L.) Merr.), small black bean (Rhynchosia nulubilis) and kidney bean (Phaseolus vulgaris var. humilis Alef.). In the study of oviposition preference, the numbers of eggs per leguminous seed on red bean, black soybean, soybean, seoritae and small black bean were 1.23, 0.61, 0.69, 1.05 and 1.13, respectively. The maximum daily number of eggs was observed at 48 hours and the minimum was at 96 hours. According to each host leguminous seed, developmental time for each host seed was different. The shortest adult emergence time was on red bean (25.27 days). The other five leguminous seeds increased or doubled the adult emergence time. Adult emergence rates feeding on red bean, seoritae, black soybean, soybean, small black bean were 83.33%, 28.23%, 27.87%, 20.44%, and 11.59%, respectively. Emergence rate on red bean was four times higher than the rate on other seeds. The longevity of emerged female adults was almost all longer than that of males. The male adults weighed the lowest of feeding on small black bean. Female adults weighed the lowest of feeding on soybean. Adult weights were the heaviest for both males and females feeding on red bean. As a result, hosts of azuki bean weevil could decrease oviposition rate, emergence rate, adult longevity, and adult weight but increase emergence time. Especially in kidney bean, adult was not completely emerged. No eggs were laid. These results suggest that there might be emergence inhibitors in kidney bean. These imformation might be used to control damages caused by azuki bean weevils.
In order to observe the growth and development of Fibricola seoulensis metacercariae, the tadpoles of Rana nigromaculata were experimentally infected with the cercariae. The meta cercariae of various developmental stages were recovered from the tadpoles after 2 to 65 days of infection. They were prepared for morphological observation, and were given orally to mice to observe their infectivity. The following results were obtained. 1. All of the tadpoles exposed to the cercariae were observed to harbour the larvae in their abdominal cavity. 2. The young metacercariae of 2 days after infection were $121.1{\mu}m$ long and $63.3{\mu}m$ wide. They grew linearly for the first 14 days to be $262.0{\mu}m$ long and $166.4{\mu}m$ wide. Thereafter, no more growth recognized until 65 days. 3. The larvae of 2 days old were similar with cercarial body and had 2 suckers, a pharynx, 2 ceca and a primordium of germ cells but no tribocytic organ. On the 8th day, they had tribocytic organ, and their morphology resembled that of mature metacercariae. 4. The metacercariae younger than 10 days could not infect the mice. Only the metacercariae older than 14 days had infectivity. The recovery rates increased by the age of metacercariae from 19.0% in 14 days old to 70.0% in 40 days old. Above findings indicate that the tadpole is indispensable for metacercarial development and it needs at least 2 weeks for maturation. The tadpole is a pivotal host in the life cycle of F. seoulensis for connection between the snail and the frog.
During the period from April to October, 1998 the authors investigated the gall development process of Chinese gallnuts (Schelchtendalia chinensis) on sumac (Rhus javanica) at Mt. Goduk near Chonju city in Chonbuk province. The majority (56.8%) of chinese gallnuts were formed on first pinnates and 24.3% on second pinnates of sumac leaves. Galls began to form from the 1st of June and grew slowly until the 24th of June. Galls enlarged rapidly after the 1st of August, but their growth rates decreased after the 26th of August and stopped growing after the 8th of September. Alate viviparous females escaped from galls after the 24th of September. Fundatricies, the second generation in the galls, exuviated three times to become adults on the 24th of June. Fundatrix adults bore 1∼10 eggs in the body and laid 1∼16 nymphs of the second generation. Nymphs of the second generation began to appear on the 17th of June and had matured completely on the 14th∼24th of July. Adults of the second generation bore eggs in the body. Thus the developmental period of the second generation was 27∼37 days. Nymphs of the third generation appeared on the 14th of July and finally alate viviparous females were developed on the 24th of September. During the two months from the beginning of the third generation to the appearence of the final alate viviparous females, there should be at least two generations, that is, the third and fourth generations. The fourth generation seems to begin at about the 20th August because the number of inhabitants (2,859 individuals) in galls on the 26 th of August had increased more than 10 times those (263 individuals) on the 19 th of August. The tannin content of galls was 65.04∼68.23% while that of sumac leaves and stems was 11.56%, 3.49% respectively.
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