• Title/Summary/Keyword: Male fertility

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A Study on the Morphological Analysis of Sperm (정자의 형태학적 특성 분석에 관한 연구)

  • Paick, Jae-Seung;Jeon, Seong-Soo;Kim, Soo-Woong;Yi, Won-Jin;Park, Kwang-Suk
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.153-165
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    • 1997
  • In male reproducible health, fertility and IVF (in-vitro fertilization), semen analysis has been most important. Semen analysis can be divided into concentration, motional and morphological analysis of sperm. The existing method which was developed earlier to analyze semen concentrated on the sperm motility analysis. To provide more useful and precise solutions for clinical problems such as infertility, semen analysis must include sperm morphological analysis. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and difficult to reproduce. Therefore, with the help of development of microcomputers and image processing techniques, we developed a new sperm morphology analyzer to overcome these problems. In this study the agreement on percent normal morphology was studied between different observers and a computerized sperm morphology analyzer on a slide-by-slide basis using strict criteria. Slides from 30 different patients from the SNUH andrology laboratory were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded. The ability of sperm morphology analyzer to repeat the same reading for normal and abnormal cells was studied. The results showed that there was no significant bias between two experienced observers. The limits of agreement were 4.1%${\sim}$-3.8%. The Pearson correlation coefficient between readers was 0.79. Between the manual and sperm morphology analyzer, the same findings were reported. In this experiments the slides were stained by two different methods, PAP and Diff-Quik staining methods. The limits of agreement were 7.2%${\sim}$-5.7% and 6.0%${\sim}$-6.3%, respectively. The Pearson correlation coefficients ware 0.76 and 0.91, respectively. The limits of agreement was tighter below 20% normal forms. In the experiments of repeatability, 52 cells stained by PAP and Diff-Quik staining methods were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs were 0.76, 0.81, 0.86, and 0.75, 0.88, 0.88 respectively. In this study it was shown that there was good agreement between manual and computerized assessment of normal and abnormal cells. The repeatability and agreement per slide of computerized sperm morphology analyzer was excellent. The computer's ability to classify normal morphology per slide is promising. Based on results obtained, this system can be of clinical value both in andrology laboratories and IVF units.

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Identification of genes involved in inbreeding depression of reproduction in Langshan chickens

  • Xue, Qian;Li, Guohui;Cao, Yuxia;Yin, Jianmei;Zhu, Yunfen;Zhang, Huiyong;Zhou, Chenghao;Shen, Haiyu;Dou, Xinhong;Su, Yijun;Wang, Kehua;Zou, Jianmin;Han, Wei
    • Animal Bioscience
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    • v.34 no.6
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    • pp.975-984
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    • 2021
  • Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.

A Study on Family Planning Rumors & Practice in Rural Communities (일 농촌지역 가족계획 풍문과 실천에 관한 연구)

  • 추수경
    • Journal of Korean Academy of Nursing
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    • v.8 no.2
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    • pp.63-77
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    • 1978
  • Studies pertaining to influential factors on adoption and practice of family planning are very much needed to lower fertility, for family planning has regarded as only one social acceptable means to curb the accelerated population growth. One of the influential factors is known as rumors on family planning methods that hinder the actual practice of family planning. This study generally aimed at disclosing rumors on family planning methods prevalent in rural communities so that one could gain some in sights to cope with the ill effects of rumors to promote family planning Practice in rural communities in Korea. In order to accomplish the general aims this study formulated following four specific objectives; 1. Find out types and frequencies of rumors exposed of contraceptive methods. 2. Find out the relationship between the rumors exposed and socio-economic and demographic characteristics of new village leaders and health workers. 3. Examine the relationship between family planing rumors and family planning practices. The materials for this study were drawn from the two different sets of data gathered in July and Octorber 1977 respectivily by Center for Population & family Planning, Yon sei University. One is current family planning practice among eligible women reaiding in 4 Myuns in Kwang Wha country and the other is Survey on Rumor on family planning method heard by new village leaders and health workers in the four myuns. The four Myuns were divided into 60 small areas. Current family planning practice rate in each small area and as a whole were calculated. The unit of analysis in this study was not the indviduel person but the 55 small areas. Percentage, average, F-Test t-test and a coefficient of correlation were used for statistical analysis. Results of this study could be summarized as follows: 1. Rumors by different method of contraception: Medical complications, gastro-intestinal disorders, and difficulty in usage one most prevalent rumors about oral pills. Around 49% -77% of the 55 areas were often heard and the most frequent rumor was related medical complications of oral pills. Rumors on medical complications and incomplete effect of intrauterines as contraceptive were heard 51-66% of the 55 small areas. Rumors that vasectomy resulted in family problems, for instance infiedelity of spouse were often heard to 44% of the 55 small areas. 2. Rumors by socio-economic and demographic characteristic of new village leader and health workers: Among the demographic characteristics such as sex, age and sex composition of children, sex and age differentials in frequency of rumors heard more observed, Female new village health workers have heard more frequently than male new village leader. (t = 7.137, p> 0.01, d.f. = 53, a coefficient of correlation 0.27, p>0.05) The Younger age group less than 40 years of age have heard rumors of than the group over 40 years old. (t = 7.18, p>0.01, d.f. = 53, a coefficient of correlation 0.27, p>0.05) However, it was not observable that a consistent sex and age differentials in frequency of rumors heard of each contraceptive mettled, But, more female new village health works heard of rumors about intrauterine device than male new village leaders. (t = 0.497, p> 0.01, d.f. = 53, a coefficient of correlatin 0.32, p>0.01) 3. It was found that there was no statistically significant relationship between the frequency of rumors heard and current family planning practices on the whole, However, frequency of rumors heard about vasectomy appeared to have a positive relationship with current practice of vasectomy. A rather consistent pattern of relationship between“requency of rumors heard”and current family practice rate was demonstrated of 10 graphes which showed the relation between the two variagles. The current family planning practice rate in the“never heard”group and “frequently heard”group was equally lower than that in“often heard”group. The relatively consistent ∩ pattern of relationship needs to be farther investigated, for this pattern is different from the relationship that has been assumed to exist between these two variables.

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STUDIES ON THE DIMORPHISM AND TRANSITION OF BISEXUALITY OF HETEROSTYLOUS POLYGONACEAE (여뀌과 이형경식물의 Dinorphism과 Bisexuality의 변화)

  • Harn, Chang-Yawl
    • Journal of Plant Biology
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    • v.3 no.2
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    • pp.6-18
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    • 1960
  • The present experiments were designed in order to clarify the differences between the long and short styled plants and the transgressive gradition in the degree of dimorphism among the three heterostylous species of the Polygonus, P. japonica, F. esculentum, and P. senticosa, based on investigations regarding the floral structure, ecological and physiological traits, the results of which are summarized as follows: (1) P. japonica, although it exhibits typical dimorphism, has undergone so high a differentiation between long and short styled that its long styled individuals behave as if they were female; and short styled individuals as if male. In long-styled individuals, filament, anther, and pollen grains show signs of degeneration, most of the pollen being abortive. On the other hand, in short styled individuals, the filament, anther, and pollen grains have attained remarkable development; the pollen grians are large and fertile. In short-plant the fertilized flowers readily drop off in every stage of their embryo development. This species has completely lost the self-fertile property, which is characteristic of the non-dimorphic Polygonum genus. Although this specsei typically exhibits the physiological characteristics of the non-dimorphic Polygonum genus. Although this specisei typically exhibits the physiological characteristics of dimorphism in controlled pollination, the short-styled individuals bear no seed in nature, thus misleading taxonomists to idenfity the short-styled plant as male. 2) The morphological feature of the flower organ of P. senticosa obviously indicates definite dimorphism. Physiologically, however, no differentiation towards dimorphism was observed, the species still retaining, both in long and short-individuals, the self-fertile property common to the Polygonum genus. Elaborate examinations revealed that regardless of the modes of pollination, both fertiization and seed setting flourish, no differentiation betwen legitimate and illegitimate unions being recognizable. This sort of physiological property has not been observed in the investigations of other heterostylous plants. It is assumed that this species is differentiated structurally into dimorphism, but not yet physiologically. In nature, however, this plant would have more opportunities to be cross-pollinated, i.e., legitimately combined, than self-pollinated because of the development of two forms of flowers. 3) In terms of heterostylism, the F. esculentum just occupies the intermediate position between P. japonica and P. senticosa structurally, ecologically, and physiologically. Doescription of some of the physiological behavior of the plant will suffice to demonstrate the above facts. While P. japonica has completely lost its self-fertile property, P. senticosa still retains it wolly. In F. esculentum 2-6% of self-fertility is the result in illegitimate combination. There occur occasionally hereditary self fertile individuals among some of the F. or 20 min. irradiation plot, when they reach any stage of the same bacterial population. In addition to this increase of total population in the plots with the more dose of UV light irradiation, it seems that the more dose of UV light irradiation is the more shortened the generation time of Azotobacter. Therefore, it is clear that variation of reproductive rate must be, mere or less, due to the genetic effects induced by UV light irradiation. On the other hand, the lag phase or logarithmic growth phase in nonirradiated culture is shortened prominently, and this must be due to the difference in bacterial number of the original inoculm. The generation time of Azotobacter is shortened by exogeneous treatment of nuclei acid derivatives, and the degree is greater in case of DNA derivatives than RNA dervatives. W.H. Price reported that the rate of ribose nucleic acid to protein in Staphylococcus muscae is proportional to the generation time: that is the faster the cell can form ribose nucleic acid, the more rapid its growth. This explains the shortening of generation time by exogeneous RNA derivatives in this work reasonably. On the other hand, it is well known that the desoxyribose nuclic acid content per cell is constant and independent of the generation time. A.D. Laren and W.N. Takahashi reported that the infectious RNA from TMV is 6 times as sensitive to inactivation by UV as it is in the form of intact virus, and that inactivation of infectious TMV involves onlu a local change on RNA chain. But, the effect of exogeneous DNA in this work suggests that irradiated living cell which cotain DNA bring about some change on DNA moleculs as well as RNA molecules. And if the mutagenic effects of UV take into consideration, it is very reasonable. Therefore, it is clear that the variation of the generation time by UV irradiation is, more or less, due to the genetic effects. Therefore, it seems that the shortness of the average lifewpan of Azotobacter by UV irradiation is resulted not only from the influence of the environmental conditions, but also from the variation of genetic factor of the individual.

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Fertility Restoration of F1 OA Interspecific Hybrid by Spontaneous Meiotic Polyploidization and Analysis of Their Progenies (자발적 생식세포 염색체 배수화 기작에 의한 Oriental-Asiatic hybrid(OA) F1 종간잡종의 임성회복과 후대검정)

  • Chung, Mi-Young;Chung, Jae-Dong;Van Tuyl, Jaap M.;Lim, Ki-Byung
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.213-219
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    • 2009
  • This study was carried out to establish breeding system for interspecific hybridization of Lilium using spontaneous meiotic polyploidization of $F_1$ OA hybrid and its progenies. Pollen viability and germination of $F_1$ OA interspecific hybrid showed variation by collection time, indicating that pollen grain in June, which was natural blooming season, had better viability and germination ability than in October. 2n-gametes from spontaneous meiotic polyploidization of OA hybrid were crossed with either diploid Asiatic or Oriental hybrid, and subsequent progenies crossed with Asiatic hybrid as female and OA hybrid as male showed the highest efficiency, where 2x-2x A$\times$OA produced 0.64 plants per cross combination, while 2x-2x, O$\times$OA and 2x-2x, OA$\times$A crosses produced 0.42 and 0.4 plant per ovary, and especially no progeny was obtained in 2x-2x, OA$\times$O cross combination. For the subsequent progeny production, a cross between diploid Asiatic hybrid as female and diploid OA hybrid as male was the best ploidy combination. Ploidy level of subsequent progeny derived from backcross with 2n-gametes of OA hybrid revealed that triploids from 2x-2x OA$\times$A, tetraploids from both 4x-2x, AA$\times$OA and OO$\times$OA, diploid from 2x-2x, O$\times$OA, and diploids or triploids from 2x-2x, A$\times$OA crosses were produced respectively.

Comparison of Attitudes toward Children among Mothers of Young Children and Maternal Grandmothers: With Specific Focus on Ideal Number of Children, Gender Preference, Expectation Old Age Security and Positive and Negative Values of Children (유아 어머니와 외할머니 세대의 자녀관련 인식 비교: 이상적 자녀 수, 자녀의 성별에 대한 선호도, 노후부양에 대한 기대, 긍정적 자녀가치와 부정적 자녀가치를 중심으로)

  • Young-Shin Park ;Uichol Kim ;Mi-Sook Kim
    • Korean Journal of Culture and Social Issue
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    • v.18 no.1
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    • pp.127-152
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    • 2012
  • The research investigates the attitudes toward children among mothers young children and maternal grandmothers, focusing specifically on the ideal number of children, gender preference, old age security and positive and negative values of children. A total of 442 respondents, consisting of 221 young mothers (average age of 35) and 221 maternal grandmothers (average age of 63) completed a questionnaire developed by Kim, Park and Kwon (2005). The reliability of the scales ranged from Cronbach α of .78 to .91. The results are as follows. First, as for the ideal number of children, young mothers reported that they would prefer two children, while grandmothers reported that they would prefer four children. As for the ideal number of male child, young mothers reported that they would prefer that they would prefer one son, while grandmothers reported two sons. Second, as for the gender of the child, young mothers did not show a gender preference, while grandmothers stated that at least one child should be a son, especially if it is the only child. Third, grandmothers had higher expectation of old age security in their male child than young mothers but two groups did not show any difference for the female child. Fourth, as for positive values of children, young mothers were more likely to emphasize personal aspects (i.e., psychological pleasure and happiness) and family cohesiveness, while grandmothers were more likely to emphasize social aspects (i.e., continuing the family line and old age security. As for negative values of children, young mothers were more likely to emphasize personal aspects (i.e., parental role and responsibility), while grandmothers were more likely to emphasize social aspects (i.e., family conflict and relationship). Fifth, as for factors that influenced the number of children that they decided to have, young mothers were more likely to report negative values of children (i.e., financial constraints), while grandmothers were more likely to emphasize positive values of children (i.e., continuing the family line).

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Performance and Meat Quality of Three-Crossbreed Korean Native Chickens (KNC) (한국토종닭 3원교배종의 생산성과 육질 특성)

  • Park, Mi-Na;Hong, Eui-Chul;Kang, Bo-Seok;HwangBo, Jong;Kim, Hak-Kyu
    • Korean Journal of Poultry Science
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    • v.38 no.4
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    • pp.293-304
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    • 2011
  • The current work was carried out to investigate the effect of crossbred Korean native chickens (KNC) on performance and meat quality. A total of 720 chicks (1d of age) was used in this work and were divided into groups by crossbreds (A, B, C and D) and sex (male and female). Crossbreds were A) (KNC egg-meat type C strains ${\times}$ KNC meat type S strains) ${\times}$ Ross broiler, B) (KNC egg-meat type C strains ${\times}$ KNC meat type H strains) ${\times}$ KNC meat type S strains, C) (KNC native R strains${\times}$KNC meat type S strains)${\times}$KNC meat type H strains and D(White Semibroiler Chickens). Experimental diets consisted of 3 phases such as starter (0~5 weeks; CP 20.0%, ME 3,050 kcal/kg), earlier (5~8 weeks; CP 18.0%, ME 3,100 kcal/kg) and finisher (8~12 weeks; CP 16.0%, ME 3,150 kcal/kg). They were fed the broiler diets for 12 weeks at the flat house and thirty six chickens were slaughtered at week 5 and 10. There was no significant difference on the fertility of crossbred KNCs, and the hatchability of B crossbred was low compared to other crossbreds. On body weight (BW), D crossbred resulted in a higher BW after 5 weeks (P<0.05). Body weight gain (BWG) and feed intake (FI) of A crossbred were also significant higher compared to the other crossbreds for all periods. On carcass ratio (CR), A crossbred showed higher CR at 5 weeks, there was no significant among crossbreds at 10 weeks. The partial meat ratio(ala, breast, neck, leg) of C crossbred was the lowest among other crossbreds (P<0.05), but the back meat ratio was not difference among treatments at 5 and 10 weeks. pH of meats have no difference among crossbreds at 5 week, and A crossbred was high compared to other crossbreds at 10 week (P<0.05). The moisture content of D crossbred meat and the protein content of B crossbred meat were high compared to other crossbreds at 5 week. Chemical compositions of meats have no difference among crossbreds at 10 week. Lightness and redness have not significance among crossbreds at 5 week, and redness of A crossbred was the highest among all crossbreds (P<0.05). Shear force and cooking loss of A crossbred were high at 5 week, and cooking loss of A crossbred was low compared with other crossbreds at 10 week. These results suggested the basic data that needed to develope the new strains.

Production of Transgenic Pigs with an Introduced Missense Mutation of the Bone Morphogenetic Protein Receptor Type IB Gene Related to Prolificacy

  • Zhao, Xueyan;Yang, Qiang;Zhao, Kewei;Jiang, Chao;Ren, Dongren;Xu, Pan;He, Xiaofang;Liao, Rongrong;Jiang, Kai;Ma, Junwu;Xiao, Shijun;Ren, Jun;Xing, Yuyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.7
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    • pp.925-937
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    • 2016
  • In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

Spermatozoa Characteristics of Streptozotocin-induced Diabetic Wistar Rat: Acrosome Reaction and Spermatozoa Concentration (Streptozotocin으로 유발된 당뇨병성 Wistar Rat 정자의 첨체반응 및 수 변화 특성)

  • Cheon, Yong-Pil;Kim, Chung-Hoon;Kang, Byung-Moon;Chang, Yoon-Seok;Nam, Joo-Hyun;Kim, Young-Soo;Gye, Myung-Chan;Kim, Moon-Kyoo;Kim, Kil-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.89-96
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    • 1999
  • Some of the information concerning sexual function in the male diabetes has been focused upon the problems of endocrine or semen parameters. However, the characteristics of acrosome reaction and spermatozoa concentration at the epididymis and vas deferens have scarcely been studied, and the causes of the infertility has not been critically identified. So, we designed to inspect the spermatozoa concentration and the characteristics of acrosome reaction at epididymis and vas deferens of diabetic Wistar rat induced by streptozotocin (STZ, 70 mg/kg, ip). Experimental animal was sacrificed at 3 days and 14 days after the STZ injection. In the diabetes-induced rat, the levels of insulin and glucose had a pattern of inverse proportion. The spermatozoa concentrations in caput and corpus epididymis were significantly decreased in all diabetic condition. In cauda epididymis, however, there was significant decrease in sperm concentration at 14 days onward. In diabetic rat, the spontaneous reaction rate of spermatozoa of cauda and vas deferens were significantly higher than the control group. The ARIC (acrosome reaction to ionophore challenge) value of caudal sperm was 28.7 at control, 22.1 at 3 days, and 8.3 at 14 days. In the present study the spermatozoa concentration was decreased and the spontaneous reaction rate was increased by diabetes. In ARIC-test, it is revealed that the fertility of spermatozoa of 14 days group was lower than control or 3 days group. Diabetes mellitus may be provoke the decreased fertilization rate and subsequent infertility and subsequent infertility.

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Overcoming sterility by Caffeine and Temperature in Oriental-Asiatic Interspecific Lily Hybrid (백합 Oriental-Asiatic 종간잡종의 임성 회복을 위한 카페인과 변온처리)

  • Park, Song Kyoung;Park, In Sook;Kim, Chang Kil;Jee, Sun Ok;Lim, Ki Byung
    • FLOWER RESEARCH JOURNAL
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    • v.17 no.4
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    • pp.279-284
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    • 2009
  • In order to recover fertility from sterile interspecific OA-1 $F_1$ hybrid (Oriental hybrid 'Mero Star' ${\times}$ Asiatic hybrid 'Connecticut King'), various concentrations (0.1, 0.3 and 0.5%) of caffeine were injected directly into flower buds and then confirmed the viability of OA-1 $F_1$ hybrid at the flowering time. After the caffeine treatment, fertilized $F_1$ hybrids were crossed as female with Asiatic hybrid 'Lanzarote' as male. Five plantlets were obtained from seven embryos of 16 pollinated flowers at 0.3% treatment of caffeine while 0.5% treatment of caffeine obtained one plant let and 0.1 % treatment of caffeine plantlet did not produce at all. Thus 0.3% of caffeine treatment was considered as optimum concentration to produce subsequent progenies and the OA-l $F_1$ hybrid treated with caffeine produced 51% of putative 2n gametes. Pollen germination of OA-2 ('Romero Star' ${\times}$ 'Lady Rosa') and OA-3 ('Expression' ${\times}$ 'Lady Rosa') was not differ between temperature treatment alone and in combination with caffeine and temperature treatment. In the reciprocal crosses of OA-1 and Asiatic hybrid 'Lanzarote' or Oriental hybrid 'Sorbonne', A ('Lanzarote') ${\times}$ OA-1 or OA-1 ${\times}$ A crosses showed better results than O ('Sorbonne') ${\times}$ OA-1 or OA-1 ${\times}$ a crosses in plant obtaining. All progenies obtained from A ${\times}$ OA-1 or OA-1 ${\times}$ A crosses were confirmed as triploids by GISH analysis.