This study was carried out to cryopreserve and to investigate characteristics of semen in Hanwoo. Semen was obtained from bulls selected by Daekwanryeong Branch station. Semen was collected each morning of the experiment, placed in water jacketed tubes at 37$^{\circ}C$, and trans-ported to the research laboratory within 10 minutes. Semen was extended with Egg yolk-glycerol extender to contain 50${\times}$10$^{6}$ sperm/ml. Semen was cooled over a 6h period in water jacketed tubes from about 25 to 5$^{\circ}C$, Egg yolk-glycerol extender was added in one step at 5$^{\circ}C$. Semen was aspirated into 0.5ml straws, which were sealed with powder. Egg yolk-glycerol extender, which is used in Hanwoo sperm frozen and stored, semen from 13 Hanwoo bulls collected, the postthawed percentages of motile sperm were 65.7%. In semen characteristics of Hanwoo bulls, number of bulls volume are 5.7 ml and total cell count are 975${\times}$10$^{6}$ m1 ejaculate.
Ecological and ethnomedicinal survey of plants was conducted in one hundred and twenty homesteads in Mbala, Amuda, Umuaku, and Nneato communities of Nneochi Local Governement Area, Abia State-Nigeria. A total of ninety-one medicinal plant species belonging to seventy-eight genera and forty-eight families, used in the treatment of malaria, yellow fever, fibroid, hepatitis, convulsion, hypertension, diabetes, insomnia, ulcer, rashes, low sperm count, snake bite, among others, were documented. Plant remedies were prepared mostly as infusions or decoctions from different plant parts with mainly water, and palm wine/gin sometimes. The highest number of medicinal plant species (73) was recorded in Mbala, followed by Amuda (71), Umuaku (68) and Nneato (61). Medicinal plant species diversity was highest in Amuda (Simpson 1-D=0.9621;H=3.663), followed by Umuaku (Simpson 1-D=0.9481; H=3.471), Mbala (Simpson 1-D=0.9345; H=3.341), and Nneato (Simpson 1-D=0.9307; H=3.277), respectively. Similarity in medicinal plant species was highest between Umuaku and Nneato (76.71%), followed by Amuda and Umuaku (75.95%), Mbala and Amuda (71.43%), while Mbala and Nneato had the lowest similarity (59.52%). The results of the study showed that traditional medicine is pivotal in the treatment of ailments in the study area, and that the indigenous people of Nneochi have recognized the need to conserve medicinal plants of importance ex situ within homesteads due to threats from unsustainable exploitation and deforestation.
The human semen ejaculated in a form of liquid state, coagulates immediately after ejaculation, and then liquefies again. However, the mechanisms of neither coagulation and liquefaction of semen have not been explained clearly so far, and very limited numbers of report are available, although the spermatology and andrology made rapid progress. This clinical study has been undertaken to investigate the liquefaction phenomena and practicability of the results might be applied to fertility and infertility problems. As a preliminary study, in this report the liquefaction time of various semen groups is measured and analysed. The following results are obtained: 1. An average liquefaction time of semen of a total of 60 subjects: 25 minutes. 2. An average liquefaction time of semen according to sperm count: 1) Normospermia group (20 cases): 34 minutes. 2) Oligospermia group (20 cases): 21 minutes. 3) Azoospermia group (20 cases): 20 minutes. 3. An average liquefaction time of semen according to abstinence period: 1) Less than 3 days group (30 cases): 22 minutes. 2) More then 5 days group (30 cases): 28 minutes. In conclusion: 1. The liquefaction time of semen of the normospermia group is longer than oligospermia group or azoosermia group. 2. The liquefaction time of semen may not be greatly influenced by the various factors such as abstinence period, semen volume, semen pH, age of the subjects and so on. 3. In routine semen analyses, it is recommended to begin the analysis at least 25 minutes after the ejaculation. 4. Further studies are required in conjunction with practical application of liquefaction mechanism in infertility and fertility control.
Ethane 1,2-Dimethane sulfonate(EDS), a toxin which specifically kills Leydig cells(LC), has been widely used to prepare the reversible testosterone(T) depletion rat model. Previous studies including our own clearly demonstrated that the dramatic weight loss of the T-dependent accessory sex organs such as epididymis and seminal vesicle in this 'LC knock-out' rats. These weight loss could be derived from massive and abrupt death of the cells via apoptotic process. The present study was performed to test the effect of EDS administration on the expression of some apoptotic genes in the rat epididymis. Adult male Sprague-Dawley rats($300{\sim}350$ g B.W.) were injected with single dose of EDS(75 mg/kg, i.p.) and sacrificed on Weeks 0, 1, 2, 3, 4, 5, 6 and 7. Tissue weights and the numbers of the epididymal sperm were measured. The transcriptional activities of the bcl-2, bax, Fas and Fas ligand(Fas-L) were evaluated by semi-quantitative RT-PCR. As expected, the weights and the sperm counts of epididymis declined progressively after the EDS treatment during Week 1 and 2. These decrements were discontinued with a gradual return towards normal during Weeks $5{\sim}7$, although the maximal recoveries of the epididymal weights(71%) and sperm count(38%) were subnormal on Week 7. The initial level of bcl-2 transcripts persisted to Week 6 then elevated significantly on Week 7. The level of bax transcripts significantly decreased on Week 6, and no remarkable change was found in the rest of the experimental period. The transcripts for the Fas in epididymis elevated during Weeks $1{\sim}2$, returned to normal on Week 3, and the level persisted to the Week 7. Similarly, the level of Fas-L transcripts elevated during Weeks $1{\sim}3$ and returned to normal after Week 4. Our results demonstrated the transient T depletion by EDS administration could induce the changes in expression of the apoptotic genes in rat epididymis. The activation of Fas and Fas-L in the epididymis of EDS-treated rats might be responsible for the initial apototic process and consequently the tissue damage and the sperm loss. Future studies will attempt to determine the precise molecular mechanism(s) of apoptosis in the rat epididymis.
Park, Se-Pill;Kim, Eun-Young;Lee, Keum-Si;Lee, Young-Jae;Shin, Hyun-Ah;Min, Hyun-Jung;Lee, Hoon-Taek;Chung, Kil-Saeng;Lim, Jin-Ho
Clinical and Experimental Reproductive Medicine
/
v.29
no.2
/
pp.129-138
/
2002
Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.
Clomiphene citrate. antiestrogen, was given to 39 infertile males whose spermatogenesis were disturbed and the efficacy of the drug was evaluated at the Department of Urology in 1980. (Table 1). Patients were divided into 3 clinical observation groups such as group I composed of 19 cases of idiopathic azoospermia, group II consisted of 15 cases of oligospermia following the vasovasostomy, and group III comprised 5 cases of testicular azoospermia. (Table 2). Clinical characteristics of these patients were as follows: Age of the patients ranged from 26 to 43 years old with mean of 34, and that of their wives ranged from 24 to 41 years old with mean of 31. Duration of marital life ranged from 1 to 21 years with mean of 5 years. Sizes of testis ranged from 6 to 25 ml with mean of 16 ml. Coital frequency ranged from 0.5 to 6 per week with mean of 2.4 per week. Levels of plasma FSH ranged from 3.15 to 23.06 lU/1 with mean of 8.15 lU/1, those of LH ranged from 2.98 to 19.89 lU/1 with mean of 8.18 lU/1 and those of testosterone ranged from 3.09 to 9.97 ng/ml with mean of 6.48 ng/ml. (Table 3). Clomiphene citrate was given in dosage of 50 mg per day (in d.) orally to 31 patients for 3 to 9 months and in dosage of 100 mg per day (b.i.d.) orally to 8 patients for 3 to 9 months. (Table 8). Semen samples were analysed monthly on each patient by routine analysis techniques. For the assessment of the efficacy of Clomiphene citrate on faulty spermatogenesis following empirical criteria were used: For semen quality: Improvement (I) represents that semen parameter increased more than 25% from basal level after the treatment, Unchange (U) expresses that semen parameter increased less than 25% of basal level or not changed after the treatment and Deterioration (D) means that semen parameter decreased from basal level after the treatment. For fertility unit (total counts ${\times}$ motility ${\times}$ morphology ${\div}10^6$): Improvement (I) represents that fertility unit increased more than 10 units after the treatment, Unchange (U) expresses that fertility unit increased less than 10 units or not changed after the treatment, and Deterioration (D) means that fertility unit decreased after the treatment. (Table 4). Results obtained from the Clomiphene therapy were as follows: Changes of spermiograme before and after the Oomiphene therapy shown in the Table 5. Sperm counts increased from 23 to 31 ${\times}10^6$/ml in group I, from 17 to 29 ${\times}10^6$/ml in group II. Other parameters of spermiogramme were not changed significantly after the treatment. Fertility units increased from 14 to 18 units after the treatment in group I, and from 16 to 18 units after the treatment in group II. Effectiveness of Clomiphene citrate on spermatogenesis was summarised in the Tables 6 and 7. After the treatment, sperm count increased in 11 patients, motility increased in 6 patients, morphology increased in 4 patients and fertility units increased in 9 patients. No sperm could be produced by Clomiphene citrate in group III of testicular azoospermia. Dosage of 50 mg of Clomiphene citrate per day for 3 to 6 months was proved to be the most effective in the present series. (Table 8). Pregnancy occurred in 2 patients after the treatment. No particular side effects were noted by the treatment. Pharmacologic compounds used for male infertility were shown in the Table 9. Reported results of Clomiphene citrate were shown in the Table 10.
This experiment was carried out to determine the optimum interval between inseminations in artifical insemination of hens. Two hundred and forty hens of Hisex commercial stock at 25 weeks of age and 20 cocks of the Rhode Island Red at 40 weeks of age were used for the experiment, and a total of 6,784 eggs were obtained. The intervals between inseminations compared in this study were: 3 days (T1), 5 days (T2), and 7 days(T3). Mixed raw semen was inseminated and the semen does was 0.03ml per insemination per hen. The inseminations were conducted at 15:00 at each time. The total number of insemination performed was 9 for the T1, 6 for the T2 and 5 for the T3, and eggs were collected over a period of 31 days, 32 days and 35 dyas, respectively. The average egg production of the hens during the experiment was 85.9% and the average temperature during the experiment was around 30$^{\circ}C$. The average sperm count was 3.69 billion per ml. The results obtained in this experiment can be summarized as follows: The fertility over the entire experimental period bythe treatment was 91.7% for the T1, 84.4% for the T2, and 75.2% for the T3. The difference between T1 and T3 in fertility was significant at 5% level. The average fertility on the second, third and fourth day after the insemination in the T2 and T3 was maintained at a relatively high level, but it tended to decline rapidly from the fifth day after the insemination. The average fertility for one week after the last insemination was 88.8% for the T1, 88.8% for the T2 and 78.6% for the T3, and none of the differences among the treatments were statistically significant. On the basis of the results from this study, it is recommended to adjust the insemination intervals within the range from the 3 to 5 days in order to maintain a highest level of fertility in the hens at an early stage of egg production as in the case of the hens used in this experiment. An insemination interval of 3 days is recommended, especially at an initial stage of insemination. For the hens with a low fertility, shortening, of the insemination interval to 3 or 2 days is desirable.
Kim, Kyung-Tae;Kim, Tae-Hong;Joo, Young-Min;Choe, Jin-Ho;Lee, Joong-Shik;Seo, Ju-Tae
Clinical and Experimental Reproductive Medicine
/
v.35
no.4
/
pp.303-308
/
2008
Purpose: The purpose of this study is to determine the effect of preoperative semen parameters on both seminal improvement and pregnancy rates following varicocelectomy. Methods: This survey was done in 278 patients who underwent microsurgical inguinal varicocelectomy from January 2001 until October 2006. By the total motile sperm counts (TMSC) before operation, the patients were stratified into three groups. Group A (mild oligoasthenospermia) was defined as above 20 million, group B (moderate oligoasthenospermia) was defined as between 5 and 20 million, and group C (severe oligoasthenospermia) was defined as below 5 million. Improvement rates of TMSC and pregnancy rates following varicocelectomy of each groups were compared. Results: The average TMSC of all the patients was 25.75 million before operation and after operation, it was 80.24 million, showing an average increase of 54.49 million (211.6%). To take a look at mean absolute increase (mean relative increase proportion), group A showed 67.90 million (131.2%), group B 62.20 million (482.5%) and group C 26.33 million (1841.2%). The patients with varicocele whose semen parameter is in bad condition show relatively a low mean absolute increase but high mean relative increase proportion. There was no significant difference in natural pregnancy rate among each groups (p=0.119, p=0.059). Conclusions: Even in the varicocele patient whose semen parameter was in bad condition before surgical operation. varicocelectomy could be chosen as the first treatment to male infertility.
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