• Title/Summary/Keyword: amount of semen

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Effect of Dimethylformamide on Post-Thaw Motility, Acrosome Integrity, and DNA Structure of Frozen Boar Sperm

  • Hwang, You-Jin;Yang, Jae-Hun;Kim, Sang-Ok;Kim, Bo-Kyung;Choi, Seon-Kyu;Park, Choon-Keun;Kim, Dae-Young
    • Journal of Embryo Transfer
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    • v.24 no.4
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    • pp.275-279
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    • 2009
  • The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

Distribution of Foreign Mineral Materials and Heavy Metals Contained in Herbal Medicines, and Effect of Washing (한약재에 함유된 광물성이물과 중금속의 분포 및 수세효과)

  • Kim, Dong-Gyu;Kim, Kyung-Sik;Lee, Sung-Deuk;Jo, Sung-Ae;Lee, Hyun-Kyoung;Jung, Sun-Ok;Jung, Kweon;Park, Seung-Kook
    • Korean Journal of Pharmacognosy
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    • v.45 no.1
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    • pp.41-47
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    • 2014
  • This study was conducted to investigate the content of foreign mineral materials and heavy metals (Pb, As, Cd, Hg) contained in commercial herbal medicines (1504 samples) classified by parts used, and the effect of removing heavy metals by washing treatment (take 50 g of the sample and put 1000 ml of distilled water, shake it gentle by hand for 1 min). The 5% trimmed means of acid-insoluble ash value (%) were as follows; above ground parts (0.76) and underground parts (0.52). Those of the total amount of individual heavy metals value (mg/kg) were as follows; caulis (1.33), flos (1.23), herba (0.91), cortex (0.76), rhizoma (0.73), radix (0.67), semen (0.44) and fructus (0.39). Acid-insoluble ash content was correlated with Pb in herbal medicines of underground parts (r=0.446) (p<0.01). After washing, the removal rate of heavy metals contained in 6 radix herbal medicines shows 33-13% respectively.

In vitro fertilization using sex-sorted boar sperm mediated by magnetic nanoparticles

  • Chung, Hakjae;Baek, Sunyoung;Sa, Soojin;Kim, Youngshin;Hong, Joonki;Cho, Eunseok;Lee, Jihwan;Ha, Seungmin;Son, Jungho;Lee, Seunghwan;Choi, Inchul;Kim, Kyungwoon
    • Korean Journal of Agricultural Science
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    • v.47 no.4
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    • pp.979-985
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    • 2020
  • A wide range of techniques have been developed to separate X or Y- chromosome-bearing sperm. In particular, bovine semen sex-sorted by using flow cytometry based on differences in the amount of DNA between X and Y chromosome bearing sperm is used in dairy farms. The first piglets were produced using sex-sorted sperm 30 years ago. However, sexed sperm have not been commercially available in pigs because the flow cytometry technique is not capable of sorting the high number of sperm required for porcine artificial insemination (AI), and the prolonged exposure to an electrical filed might damage to the DNA in sperm. The purpose of this study was to evaluate a boar sperm sorting method based on magnetic nanoparticles. A flow cytometer assay verified the efficacy of the magnetic nanoparticles (> 90% of sex-sorted sperm). In addition, a duplex polymerase chain reaction (PCR) assay using sex chromosome specific genes including SRY (sex-determining region Y; male), ZFY (zinc finger protein Y-linked; male), and ZFX (zinc finger protein X-linked; female) showed that in vitro fertilized porcine embryos by X and Y-chromosome bearing sperm were 100% female (40/40) and 72% female (35/48), respectively, at 8-cell or morula stages, suggesting that the sex-sorted sperm were fertile. In conclusion, our findings suggest that the sex-sorted method based on magnetic nanoparticles can be utilized for porcine sex-sorted AI.

A Study on Abdominal Temperature of Male Infertility Patients (남성 불임 환자의 하복부 온도에 관한 임상적 고찰)

  • Lee, Chang-Hoon;Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub;Wee, Hyo-Sun
    • Journal of Oriental Medical Thermology
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    • v.4 no.1
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    • pp.23-28
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    • 2005
  • Purpose : To examine the relation between male infertility and cold hypersensitivity on lower abdomen. Methods : From 2004, 4 to 2005, 10, a total 60(infertile male 30, normal male on semen analysis 30) patients who visited the Oriental gynecological department of Conmaul Oriental Medicine hospital was selected, and their medical records were reviewed retrospectively. We compared the temperature of 3 points(CV17, CV12, CV4) of both group by DITI(Dorex, DITI-16UTI and DITI-Spectrum 9000 MB). Results and Conclusion : The general characteristics such as age, weight, height, BMI, amount and frequency of drinking and smoking of both group were not different statistically. The mean ${\Delta}T1$ between upper and lower abdomen in infertile group was higer than in normal group but they were not different statistically. The mean ${\Delta}T2$ between chest and lower abdomen was $0.46{\pm}0.41$ in infertile group, and $0.18{\pm}0.56$ in normal group, and it shows significant difference statistically. There was no significant relationship between sperm motility and ${\Delta}T1$, ${\Delta}T2$ in infertile group. In this study, we suggest that an inclination of cold hypersensitivity of lower abdomen of infertile male, but it is expressed when it is contrasted to chest of the body rather than upper abdomen.

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Effects of Aqueous Extracts from Twelve Herbs on ${\alpha}$-melanocyte Stimulating Hormone-induced Melanogenesis in B16F10 Mouse Melanoma Cell (한약재 12종의 열수추출물이 ${\alpha}$-melanocyte stimulating hormone에 의해 유도된 B16F10 흑색종 세포의 멜라닌형성에 미치는 영향)

  • Lee, Soo-jin;Choi, Yung-Hyun;Lee, Yong-Tae;Choi, Byung-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.5
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    • pp.1271-1274
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    • 2006
  • We investigated the effects of aqueous extracts from twelve medical herbs on ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis in B16F10 mouse melanoma cell. The cells were incubated with ${\alpha}$-MSH and aqueous extracts 5 days and 2 days and then analysed melanin amount and tyrosinase activities, respectively. Nine aqueous extract of herbs examined at 1 mg/m${\ell}$ level decreased melanin synthesis in B16F10 cell, especially in Agastache rugosa, Leonurus siviricus and Murus bombycis. The significant decrease of released extracellular melanin were also observed in treated cells with aqueous extract from Leonurus siviricus, Murus bombycis and Ledebouriella seseioides. The ${\alpha}$-MSH-induced activation of tyrosinase was inhibited in cells treated with aqueous extract from Cuscutae semen, Angelica tenuissima and Agastache rugosa. These results suggest that herbs inhibiting melanogenesis through tyrosinase activity may apply to develop whitening drugs and cosmetics.

Sperm DNA fragmentation and sex chromosome aneuploidy after swim-up versus density gradient centrifugation

  • Kim, Sung Woo;Jee, Byung Chul;Kim, Seul Ki;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.4
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    • pp.201-206
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    • 2017
  • Objective: The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount of sperm with fragmented DNA, sex chromosome aneuploidy, and abnormal chromatin structure. Methods: Semen samples were obtained from 18 healthy male partners who attended infertility clinics for infertility investigations and were processed with swim-up and DGC. The percentages of sperm cells with fragmented DNA measured by the sperm chromatin dispersion test, normal sex chromosomes assessed by fluorescence in situ hybridization, and abnormal chromatin structure identified by toluidine blue staining were examined. Results: The percentage of sperm cells with fragmented DNA was significantly lower in the swim-up fraction (9.7%, p= 0.001) than in the unprocessed fraction (27.0%), but not in the DGC fraction (27.8%, p= 0.098). The percentage of sperm cells with normal X or Y chromosomes was comparable in the three fractions. The percentage of sperm cells with abnormal chromatin structure significantly decreased after DGC (from 15.7% to 10.3%, p= 0.002). The swim-up method also tended to reduce the percentage of sperm cells with abnormal chromatin structure, but the difference was not significant (from 15.7% to 11.6%, p= 0.316). Conclusion: The swim-up method is superior for enriching genetically competent sperm.

Quantitative Analysis of the Marker Constituents in Yongdamsagan-Tang using Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (LC-ESI-MS/MS를 이용한 용담사간탕의 주요 성분 분석)

  • Seo, Chang-Seob;Ha, Hyekyung
    • Korean Journal of Pharmacognosy
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    • v.48 no.4
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    • pp.320-328
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    • 2017
  • Yongdamsagan-tang has been used to treat the urinary disorders, acute- and chronic-urethritis, and cystitis in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous analysis of the 20 bioactive marker compounds, geniposidic acid, chlorogenic acid, geniposide, liquiritin apioside, acteoside, calceolarioside B, liquiritin, nodakenin, baicalin, liquiritigenin, wogonoside, baicalein, glycyrrhizin, wogonin, glycyrrhizin, wogonin, saikosaponin A, decursin, decursinol angelate, alisol B, alisol B acetate, and pachymic acid in traditional herbal formula, Yongdamsagan-tang. Chromatographic separations of all marker compounds were conducted using a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization source in the positive and negative modes. The flow rate was 0.3 mL/min and injection volume was $2.0{\mu}L$. The correlation coefficient of 20 marker compounds in the test ranges was 0.9943-1.0000. The limits of detection and quantification values of the all marker components were 0.11-6.66 and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the optimized LC-ESI-MS/MS method, three compounds, geniposidic acid (from Plantaginis Semen), alisol B (from Alismatis Rhizoma), and pachymic acid (from Poria Sclerotium), were not detected in this sample. While the amounts of the 17 compounds except for the geniposidic acid, alisol B, and pachymic acid were $0.04-548.13{\mu}g/g$ in Yongdamsagan-tang sample. Among these compounds, baicalin, bioactive marker compound of Scutellariae Radix, was detected at the highest amount as a $548.13{\mu}g/g$.

Glucose-6-phosphate dehydrogenase deficiency does not increase the susceptibility of sperm to oxidative stress induced by H2O2

  • Roshankhah, Shiva;Rostami-Far, Zahra;Shaveisi-Zadeh, Farhad;Movafagh, Abolfazl;Bakhtiari, Mitra;Shaveisi-Zadeh, Jila
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.4
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    • pp.193-198
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    • 2016
  • Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect. G6PD plays a key role in the pentose phosphate pathway, which is a major source of nicotinamide adenine dinucleotide phosphate (NADPH). NADPH provides the reducing equivalents for oxidation-reduction reductions involved in protecting against the toxicity of reactive oxygen species such as $H_2O_2$. We hypothesized that G6PD deficiency may reduce the amount of NADPH in sperms, thereby inhibiting the detoxification of $H_2O_2$, which could potentially affect their motility and viability, resulting in an increased susceptibility to infertility. Methods: Semen samples were obtained from four males with G6PD deficiency and eight healthy males as a control. In both groups, motile sperms were isolated from the seminal fluid and incubated with 0, 10, 20, 40, 60, 80, and $120{\mu}M$ concentrations of $H_2O_2$. After 1 hour incubation at $37^{\circ}C$, sperms were evaluated for motility and viability. Results: Incubation of sperms with 10 and $20{\mu}M\;H_2O_2$ led to very little decrease in motility and viability, but motility decreased notably in both groups in 40, 60, and $80{\mu}M\;H_2O_2$, and viability decreased in both groups in 40, 60, 80, and $120{\mu}M\;H_2O_2$. However, no statistically significant differences were found between the G6PD-deficient group and controls. Conclusion: G6PD deficiency does not increase the susceptibility of sperm to oxidative stress induced by $H_2O_2$, and the reducing equivalents necessary for protection against $H_2O_2$ are most likely produced by other pathways. Therefore, G6PD deficiency cannot be considered as major risk factor for male infertility.

Inhibition Effects on Oxidative DNA Damage and Anti-inflammatory Effects of Nelumbinis Flos (연꽃의 산화적 DNA 손상 억제 활성 및 항염증 효과)

  • Jeong, Hyung Jin;Park, Yeon Gyeong;Jang, Tae Won;Kim, Do Wan;Jeong, Jin Boo;Park, Jae Ho
    • The Korea Journal of Herbology
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    • v.32 no.3
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    • pp.45-53
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    • 2017
  • Objective : Nelumbo nucifera, its rhizome and semen have been used as a traditional medicine which was studied on antioxidant, hepatoprotective effect, anti-obesity and the others. However, Nelumbinis Flos have not studied. We investigated protective effects on oxidative DNA damage and anti-inflammatory effects of Nelumbinis Flos. Methods : The antioxidant activity was conducted by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay and reducing power assay. Total phenolic content was analyzed. Also, phenolic compounds were detected by HPLC/UV. The inhibitory effect on oxidative DNA damage was determined using ${\Phi}X-174$ RF I plasmid DNA cleavage assay. The anti-inflammatory effect of Nelumbinis Flos was measured by the amount of nitric-oxide (NO) produced and protein levels of iNOS, and COX-2 in LPS induced RAW 264.7 cells. Results : The results of DPPH and ABTS radical scavenging activity at $200{\mu}g/m{\ell}$ of extraction were $97.02{\pm}0.88%$ and $96.42{\pm}0.25%$. Reducing power (fold of L-ascorbic acid as control) was $100.14{\pm}0.31$ at $200{\mu}g/m{\ell}$. Total phenol content was $8.70{\pm}0.02mg/g$. Chlorogenic acid, catechin and epicatechin were found by HPLC. Nelumbinis Flos has inhibitory effect in dose-manner against oxidative DNA damage. In addition, it showed the anti-inflammatory effect by suppression of NO production as well as protein levels of iNOS, and COX-2. Conclusion : This study suggested that Nelumbinis Flos showed potential antioxidant and suppression activities of various factors were related in NO produced. Therefore, Nelumbinis Flos as natural plant resources that may help reduce inflammation and alleviate DNA damage.

Development of High-Functional Hyaluronic Acid/Salmon Extract Formulation Using Gamma-Ray (감마선을 이용한 고기능성 히알루론산/연어추출물 제형개발)

  • Kweon, Dong-Keon;Shim, Jae-Goo;Ha, Man
    • Journal of the Korean Society of Radiology
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    • v.12 no.1
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    • pp.9-16
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    • 2018
  • Hangover after drinking is different from person to person symptoms and degree, but usually thirst, fatigue, headache, general boredom, gastrointestinal disorder, vomiting, diarrhea, deficiency of vitamin appears. This hanging phenomenon is caused by the action of precursors such as ethyl acetate and acetaldehyde, which are the by products of fermentation contained in alcohol and alcohol accumulated in hepatocytes and body. In order to solve the hangover phenomenon, the same Origin as polysaccharide Polydeoxyribonucleotide, which is a nucleic acid-sugar-phosphate complex, which is a semen or testicular extract in salmon extract, and a water soluble salmon extract powder having the same structure and lower price than Polydeoxyribonucleotide And D-Glucuronic acid and N-Acetyl glucosamine. It has excellent biocompatibility, viscoelasticity and moisturizing power. It has effect on reduction of body water loss and skin moisture content in hangover phenomenon. It is antioxidant and skin moisturizing effect Hyaluronic acid was irradiated with gamma rays, and the composition was prepared by using the salmon extract powder and the main raw material. The ethanol degradation, the acetaldehyde reduction amount, the blood acetaldehyde concentration and the acetic acid concentration were measured to evaluate the alcoholysis effect, Skin moisture evaporation rate To examine the evaluation unit water content of the skin was improved determine whether the antioxidant and provide skin moisturizing effect. The addition of ethanol extracts of salmon extracts showed a decrease of 5 to 7 times compared with no addition, and a decrease of 3 to 5 times of acetaldehyde. In addition, the change of acetaldehyde concentration and acetic acid concentration in blood showed a rapid decrease compared to the no - added control group. In addition, when the raw material of hyaluronic acid was used, skin moisture content was high and skin moisture evaporation amount was decreased. Therefore, hyaluronic acid, which is a polysaccharide polymer, has excellent viscoelasticity and moisturizing ability, It is considered to provide antioxidant and skin moisturizing effect. Therefore, it can be said that the composition containing salmon extract powder and hyaluronic acid as a main ingredient is effective for the hangover phenomenon which occurs after drinking.