This study was conducted to investigate the effects of donor cell type, individual, passage number and trypsinization time on the in vitro development of bovine somatic cell nuclear transfer embryos. Three cell types (skin, muscle and cumulus cells) and cells from 3 individuals were used for nuclear transfer. Cell were passaged by 5, 15 or 30 times, and cell were trypsinized for 1 or 3 min before injection. Nuclear transfer were performed by conventional fusion method. Development rates to the blastocyst stage were not significantly different among three cell types (16.5∼23.9%) and individuals (16.4∼19.5%). Blastocyst formation rate of cloned embryos reconstituted with cells at passage 30 (5.8%) was significantly lower than those of embryos reconstituted with 5- and 15-passaged cells (25.3 and 23.5%, respectively, P<0.05). The rate of embryos developed to the blastocyst stage was higher in embryos reconstituted with cells trypsinized for 1 min (30.7%) compared to embryos reconstituted with cells trypsinized for 3 min (P<0.05). The result of the present study indicates that different donor cell types and individuals used in this study did not affect the development of cloned bovine embryos. However, passage number and trypsinization time of donor cells affect the in vitro development of cloned bovine embryos.
In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.
Struggle refers to the process of overcoming various difficulties for a goal. The spirit of struggle is a positive attitude and reaction reflected in the process of struggle. Cultivating the spirit of struggle of college students is the call of the new era. In essence, the cultivation of the spirit of struggle is a process of learning, which is in line with Bandura's Observation Learning Theory(Bandura, 1977):Attention, Maintenance, Reproduction and Motivation. The cultivation of College Students' spirit of struggle in the new era is also a learning process of enriched experience. It is necessary to cultivate the spirit of struggle into the soul of college students and make it become a habit of students. Moreover, it is crucial to carry out adaptive transformation of Bandura's observation learning theory. By studying the mechanism of the spirit of struggle of college students, taking innovation and entrepreneurship education as a means, and aiming at cultivating the connotation of President Xi's thought on socialism with Chinese characteristics for a new era, this paper constructs the AIST model for cultivating the spirit of struggle of college students in the new era. This model includes online learning acceptance platform(Acceptance), classroom experience stimulation platform(Inspiration), iterative training solidified platform (Solidification), and competition practice transfer platform(Transfer). The purpose of this model is to provide a practical way for universities to fulfill the fundamental task of moral education and cultivate qualified socialist builders and successors. The number of students using the online learning acceptance platform ranked the first among that of the similar courses in China; The classroom experience stimulation platform and the iterative training solidified platform support each other, with an effective rate of 97%; The competition practice transfer platform has realized the continuous growth of the number of awards won in competitions for three years. The direction of future efforts is to establish the external mechanism of the spirit of struggle, to ensure the effectiveness of classroom experience and iterative training, to cultivate teachers with coaching skills, and to accurately measure the transformation point of external and endogenous motivation.
The objective of this study was to optimize the vitrification method of in vitro produced bovine embryos. Thus, in vitro produced embryos at 8 cell, morula and blastocyst stages were vitrified on electron microscope grids (EM grids) or in open pulled straws (OPS) with EG5.5 (5.5 M ethylene glycol, 1.0 M sucrose and 10% FBS in m-DPBS medium) freezing solution and their survival rates after thawing were compared. The embryos on EM grids or in OPS were briefly exposed to EG5.5 freezing solution and plunged directly into liquid nitrogen within 30 to 35 sec. Post-thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-DPBS, each for 1 min, and then cultured in CRI aa medium supplemented with 10% FBS. Embryonic survival rate was assessed as re-expanded and hatched rates of those embryos after warming. The rates of re-expansion embryos did not significantly different between EM grid (8 cell: 42.10%, morula: 66.66% and blastocyst: 77.08%) and OPS (8 cell: 47.36%, morula: 61.90% and blastocyst: 83.33%) methods. In addition, the hatched rates in EM grid (8 cell: 31.57%, morula: 57.14% and blastocyst: 72.91%) were similar to those in OPS (8 cell. 34.21%, morula: 50.00% and Blastocyst: 77.08%). Interestingly, even at the same blastocyst stage, the in vitro survival of day 7 embryos (EM grid: 79.48 and OPS: 87.18%) was higher than those of day 8 embryos (EM grid: 72.10 and OPS: 82.06%). The total cell number of blastocyst developed in vitro after vitrification was examined with Hoechst 33342 staining to compare the embryo quality among different treatment groups. The total cell number of blastocyst was not significantly different between vitrified groups (EM grid: 162.4$\pm$8.0 and OPS: 158.4$\pm$7.1) and unvitrified control (168.0$\pm$5.6). These results indicate that both vitrification containers can provide the high rate of embryo survival. Moreover, the OPS container may not need a cap to protect the container from floating after immersion in L$N_2$. Therefore, this study suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification method using EM grid or OPS with EG5.5 freezing solution. In the future, the Pregnancy rate would be investigated after transfer of our vitrified embryos into the appropriated recipients.
This experiment was conducted to study effect of cold water damage on some growth characters related to source and sink at reproductive growth stage in Jinan (sea level 303m). The cold water irrigation duration had irrigated 4, 8 and 12 days at panicle formation stage and reproduction division stage compared to perennial water irrigation. Cold water irrigation shortened culm length and panicle length and degree of panicle exsertion. The shortening effect appeared great at lower internodes when treated at panicle formation stage but at higher internodes when treated at reduction division stage. Cold water irrigation decreased the number of secondary branches and spikelets per panicle, and increased the number of degenerated spikelets being high degeneration when treated at panicle formation stage. Spikelet sterility and impediment of grain filling were affected by duration of cold water irrigation being great when treated at spikelet primodium differentiation stage and reduction division stage in particular. Grain weight was also reduced. Significant relationship existed between spikelets sterility, grain filling and yield. The degeneration of secondary branches and spikelets correlated with leaf area but spikelet sterility and yield with culm length, panicle length and panicle exsertion.
Henry Onyebuchi Okonkwo;Olubunmi Ayokunle Koyejo;Joseph Okechukwu Ariwaodo;Nsien Iniobong Bruno
Journal of Forest and Environmental Science
/
v.39
no.2
/
pp.73-80
/
2023
Little is known of the life history of Garcinia kola; the objective of this study, therefore, was to assess the fruiting age and tree size of the species in a coastal humid tropical climate plantation condition. A total 103 trees were used in the study viz; 80 ten-year-old trees at reproductive maturity onset and 13 thirty-year-old trees with several cycles of reproduction that constitute two independent variables. Data collected were age of onset of flowering and size at reproductive maturity onset. Relative size at reproductive maturity onset (RSOM) was estimated as size at reproductive maturity onset (SOM) divided by asymptotic maximal size (AMS). Data analysis was conducted using pairwise t-test and principal component analysis (PCA). Reproductive maturity onset (flowering) was recorded in the ten-year-old stand eight (8) years after planting. Mean size at reproductive maturity onset (SOM) was height 5.32±1.7 m, dbh 0.11±0.03 m, total number of branches was 29.6±7.3, crown depth 5.24±1.05 m, crown diameter was 4.78±0.7 m, branch diameter 0.098±0.01 m, leaf length 0.13±0.02 m, leaf breadth 0.37±0.01 m, twig length 0.35±0.11 m and leaf per twig 6±0.84 and asymptotic maximal size (AMS) was height 19.85±0.76 m, dbh 0.95±0.09 m, total number of branches 62±5, crown depth 18.83±0.7 m, crown diameter 12.5±1.64 m, branch diameter 0.5±1.6 m, leaf length 0.16±0.023 m, leaf breadth 0.45±0.12 m, twig length 0.37±0.11 m and leaf per twig 19±7.5. Pairwise t-test analysis showed there was significant differences between SOM and AMS in all growth factors except leaf length, leaf breadth, and twig length. Highest relative size at reproductive maturity onset (RSOM) was recorded in leaf length 0.82, twig length 0.82, and leaf breadth 0.80, while, the lowest was branch diameter 0.11. Four components out of the total of eleven were extracted to explain the relationship in RSOM: Principal component one (PC1) explained 37.23%; PC2 26.4%, PC3 22.73%, and PC4 13.64%.
Kim Y. S.;Song S. H.;Cho S. K.;Kwack D. O.;Kim C. W.;Park C. S.;Chung K. H.
Reproductive and Developmental Biology
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v.29
no.3
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pp.201-205
/
2005
The objective of this study was to investigative the effects of amino acids supplementation on maturation, fertilization and embryo development of pig oocytes. Essential amino acids (EA), non-essential amino acids (NA) or both amino acids (EA + NA) were supple-mented to North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (pFF). When the amino acids were supplemented to the maturation medium, the maturation rates were higher (p<0.05) in the NA group than control ($83.3{\pm}0.04\%\;versus\;70.0{\pm}0.05\%$, but the subsequent cleavage rates and development to morula and blstocyst stage between aminoacid supplement groups and control were not different. The developmental rates to morula and blastocysts stage were not significantly different regardless of amino acid supplementation to culture medium. In addition, supplementation of amino acids did not significantly affect the rate of fertilization and polyspermy. When the amino acids were supplement to culture medium, the number of trophectodermal (TE) cells was significantly (p<0.05) higher in amino acid supplement group than that of control ($18.6{\pm}0.5\;versus\;16.1{\pm}0.6$), whereas the numbers of inner cell mass (ICM) cells were not different among the treaonent groups and control ($29.0{\pm}0.9\~31.5{\pm}1.2$). Total cell number was also significantly (p<0.05) higher in EANA group ($50.0{\pm}1.0$) than that of control group ($44.2{\pm}1.1$). These results indicate that the amino acid supplementation to maturation and culture medium may not significantly stimulate early embryo development, but may improve the TE cell number of blastocyst stage in the pig.
Jo, Chang-Wook;Park, Cho-Rong;Yoon, Kyu-Sik;Kang, Min-A;Kwon, Hae-Ri;Kang, Eun-Jin;Seo, Mi-Ja;Yu, Yong-Man;Youn, Young-Nam
Korean journal of applied entomology
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v.48
no.3
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pp.301-310
/
2009
For the comparing of mortality of the resistance and susceptible population of Myzus persicae, etofenprox was treated in the recommended concentration of 200ppm. Mortalities of resistance population were 16.7 and 36.7%, and susceptible population were 86.7 and 86.7% after 24 and 48 hours treatment, respectively. For the detect of cross resistance to other pyrethroids, 6 pyrethroids were examined. Mortalities of susceptible and resistance populations were 90 and 31% to deltamethrin, 92 and 23% to lambda cyhalothrin, 81 and 14% to cypermethrin, 70 and 20% to $\alpha$-cypermethrin, 29 and 28% to fenpropathrin, 84 and 29% to fenvalerate, respectively. It was showed that resistance populations were generally resistive to other pyrethroids. On the other hands, for recognized ecological characteristic of M. persicae susceptible and resistance populations life table was tested on the pepper leaves in the petri dish and on the plant in the pot. This results were showed that intrinsic rate of increase ($r_m$), net reproduction number ($R_0$) and generation time in day ($T_c$) were significantly different between two population in both tested. However, life span and reproduction period were slightly different between them. Otherwise, feeding behaviors were tested using EPG technique with non- and treated etofenprox. First potential drop time of susceptible and resistance population was 73.5 and 257.9 sec with non-treated and 93.3 and 1076.2 sec after treated, respectively. Electrical probing signals were 8.2 and 48.8 times with susceptible and resistance individuals after treated etofenprox, respectively. It was supposed that the resistance is more probings than susceptible population. After treated, total feeding time have more 6,728.9 sec on resistance than 965.5 sec on susceptible population. So, total non penetration time of susceptible population was 3,000 sec longer than resistance population.
This experiment was conducted to investigate how the lactation regulation such as restricted-lactation and early weaning during the suckling period influences on ovarian functions and change in serum levels of progesterone in primiparous rats. All the rats were raised in the individual cage from a few days before parturition through the suckling period. The normal lactation(NL) groups were controled 8 pups. The restricted-lactation(RL) and weaned(W) groups were subdivided into 5 subgroups as RL0, RL5, RL10, RL15 and RL20 as well as W0, W5, W10, W15, and W20 according to the day of onset of suckling. The number of pups were regulated from 8 to 4 on experimental strating day in RL gropus, and also perfectly weaned on the each on-set day in W groups. The results obtained were summarized as follows: 1. During the whole suckling period of 25 days the pups in RL group grew significantly(P<005) faster than those in normal-lactation(NL) group. The pups in earlier RL group grew significantly(P<0.05) faster than those in later RL rats, and there was no found any significant difference in body weight of pups between RL20 and NL group. The gestation period and litter size were found to be 21.53$\pm$0.04 days and 13.75$\pm$0.07, respectively. 2. The estrous cycle was not expressed in the NL group through the whole suckling period. An irregular estrous cycle was found around day 20 in RL0 group, and the regular estrous cycles were exhibited continuously from day 10 in the day 0 weaned rats. 3. In the rats of NL group the serum progesterone concentration increased from 33.16$\pm$2.64ng/$m\ell$ on day 0 to 122.5$\pm$53.68 ng/$m\ell$ on day 10, and then decreased slightly to 97.30$\pm$3.21 ng/$m\ell$ on day 20, but then decreased abruptly. However, the serum level of progesterone decreased greatly(P<0.05) in 5 to 10 days following suckling restriction in the rats from which suckling began to be under restriction on day 0 or day 15. In the early weaning group the significant ( (P<0.05) decrease in progesterone concentration was found similarly in 48 hours following weaning in all the rats weaned on day 0 through day 20. It was suggested that lactation stimulation is a very pivotal on the function of ovary.
This study was carried out to investigate the reproductive status and the effect of progesterone treatment on the recovery of reproductive disorders in Hanwoo. Hanwoo farms were surveyed the general management status, such as the type of barn, the feed intake, the incidence of reproductive disorders. The reproductive disorder cattle were treated 7 days insertion of control internal drug releasing for cattle (CIDR) and injection of PGF$_2$$\alpha$ at 1 day before removal. The recovery of reproductive disorders was assessed by determining the pregnancy following artificial insemination. The total number of surveyed Hanwoo farms was 127, and the total incidence rate of reproductive disorders was 19.7% (209/1,061). Compared to the herd size, the incidence rates of reproductive disorders in less than 10 heads (37.5%) was significantly (P<0.05) higher than in 10 to 20 heads (14.7%) and more than 20 heads (13.6%) of herd size per farm. The incidence rate of reproductive disorders in tie stall barn was significantly (P<0.05) higher than in free stall barn (30.4% vs. 14.7%), and even in free stall barn, that tended to decrease as the floor area was larger. The incidence rate of reproductive disorder by the parity was highest in heifer (50.7%), and that tended to decrease as the parity was increased. The distribution rate of the case of reproductive disorder in anestrus, recovery rate of reproductive disorders fellowing CIDR treatment was 75.1% (157/209). In the recovery rate of reproductive disorders by body condition score (BCS), BCS 1, 2 and 3 was significantly (P<0.05) higher pregnancy rate (85.7, 84.9 and 86.8%), and gross recovery rate in emaciated cattle was better than in obese cattle (BCS 4, 5). In conclusion, the incidence rate of reproductive disorders in Hanwoo raised in Youngju province area was 19.7%, and that tended to decrease as the herd size was increase because of increased farmer's attentions, and the floor area was larger. The majority of the case of reproductive disorders was anestrus. The recovery rate of reproductive disorders following CIDR treatment was 75.1%, and to optimize the recovery rate of reproductive disorders, cows and heifers were maintained BCS 1, 2 and 3 by moderate management.
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