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2.
Int. braz. j. urol ; 47(2): 275-283, Mar.-Apr. 2021. tab, graf
Article in English | LILACS | ID: biblio-1154474

ABSTRACT

ABSTRACT Purpose: Sperm DNA fragmentation is a major cellular mechanism underlying varicocele-related male infertility. However, the type of DNA fragmentation - whether oxidative or of another nature - remains unknown. Thus, the aim of this study was to evaluate single- and double-stranded sperm DNA fragmentation, and oxidative-induced sperm DNA damage in men with varicocele. Materials and Methods: A cross-sectional study was performed, including 94 normozoospermic adults, of which 39 men without varicocele (controls) and 55 men with varicocele grades II or III, uni- or bilaterally. All men collected semen by masturbation. After semen analysis, the remaining volume was used for evaluation of three types of sperm DNA damage: (i) total DNA fragmentation, using an alkaline comet assay, (ii) double-stranded DNA fragmentation, using a neutral comet assay, and (iii) oxidative DNA damage, using an alkaline comet assay associated with the DNA glycosylase formamidopyrimidine enzyme. In each assay, percentage of sperm with any degree of DNA fragmentation, and with high DNA fragmentation were compared between the groups using an unpaired Student's t test or a Mann-Whitney test. Results: The varicocele group presented a higher rate of sperm with fragmented DNA (both any and high DNA fragmentation), considering single-stranded DNA fragmentation, double-stranded DNA fragmentation, or a combination of both, as well as oxidative- induced DNA fragmentation. Conclusions: Patients with varicocele have an increase in sperm DNA fragmentation levels, particularly in oxidative stress-induced sperm DNA damage.


Subject(s)
Humans , Male , Adult , Varicocele/genetics , Infertility, Male/genetics , Sperm Motility , Spermatozoa , Cross-Sectional Studies , Oxidative Stress , DNA Fragmentation
3.
National Journal of Andrology ; (12): 922-925, 2020.
Article in Chinese | WPRIM | ID: wpr-880293

ABSTRACT

Objective@#To explore the distribution of Traditional Chinese Medicine (TCM) syndrome types and their relationship with semen parameters in infertility male patients with varicocele (VC).@*METHODS@#Using Questionnaire on Clinical Symptoms of Varicocele-Caused Male Infertility, we made an investigation among 147 infertility male patients with VC, determined the types of their TCM syndromes, obtained their semen parameters, and analyzed the distribution of the TCM syndrome types and their correlation with semen parameters.@*RESULTS@#Of the TCM syndrome types identified, kidney deficiency and stagnated heat constituted the largest proportion (34.7%), and the mixed type accounted for a significantly higher percentage than the simple type (P < 0.05). The patients with kidney deficiency and stagnated heat, compared with those with other syndrome types, had a dramatically lower sperm concentration ([21.62 ± 9.25] vs [28.88 ± 12.92] ×10⁶/ml, P < 0.01), but a higher percentage of morphologically abnormal sperm ([98.33 ± 0.15]% vs [96.27 ± 0.18]%, P < 0.05) and DNA fragmentation index ([19.72 ± 3.17]% vs [10.96 ± 3.82]%, P < 0.01). No statistically significant differences were observed in the percentage of progressively motile sperm among different TCM syndrome types.@*CONCLUSIONS@#Kidney deficiency and stagnated heat is a main TCM syndrome type in infertility male patients with varicocele and correlated with sperm concentration, the percentage of morphologically abnormal sperm and DNA fragmentation index.


Subject(s)
DNA Fragmentation , Humans , Infertility, Male/diagnosis , Male , Medicine, Chinese Traditional , Semen , Semen Analysis , Sperm Count , Sperm Motility , Spermatozoa , Syndrome , Varicocele/diagnosis
4.
National Journal of Andrology ; (12): 900-905, 2020.
Article in Chinese | WPRIM | ID: wpr-880289

ABSTRACT

Objective@#To analyze the relationship of Mycoplasma genitalium (MG) infection with routine semen parameters and sperm DNA integrity in male infertility patients.@*METHODS@#Totally, 114 semen samples, 34 MG-positive and 80 MG-negative, were collected from male infertility patients and subjected to routine semen analysis with the computer-assisted sperm analysis system, Papanicolaou staining for observation of sperm morphology, and sperm chromatin diffusion (SCD) test for detection of sperm DNA integrity. Semen parameters and DNA integrity were compared between the MG-positive and MG-negative groups with SPSS 21.0 statistical software and the relationship between the semen parameters and DNA integrity analyzed by Pearson correlation analysis.@*RESULTS@#The MG-positive samples, compared with the MG-negative ones, showed significantly decreased semen volume ([2.87 ± 0.37] vs [3.86 ± 0.43] ml, P 0.05).@*CONCLUSIONS@#MG infection may be an important factor affecting sperm quality in male infertility patients. Active prevention and treatment of MG infection can help prevent male infertility.


Subject(s)
DNA Fragmentation , Humans , Infertility, Male/microbiology , Male , Mycoplasma Infections/complications , Mycoplasma genitalium , Semen , Semen Analysis , Sperm Count , Sperm Motility , Spermatozoa
5.
Rev. cuba. endocrinol ; 30(3): e194, sept.-dic. 2019. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1126441

ABSTRACT

RESUMEN Introducción: Se estudian los factores que afectan el ADN espermático en la actualidad y en qué medida influyen en la aplicación de las técnicas de reproducción asistida de alta tecnología. La fertilización in vitro es un tema de interés que ha motivado a muchos investigadores. Objetivo: Determinar la relación que existe entre el índice de fragmentación del ADN de la cromatina espermática y los resultados de la técnica de fertilización in vitro en parejas infértiles. Métodos: Se realizó un estudio descriptivo transversal en 107 parejas infértiles remitidas del Programa Nacional de Atención a la pareja infértil, donde se precisó la asociación entre el potencial fertilizante y las variables de resultados: tasa de fertilización, calidad embrionaria y embarazo. Se estudiaron, además, algunos factores que pueden influir en el grado de fragmentación del ADN. Resultados: El potencial fertilizante no se relacionó con la tasa de fertilización, ni con el embarazo clínico, pero sí con la calidad embrionaria (P= 0.036). La edad paterna resultó ser estadísticamente significativa en relación con el potencial fertilizante (P= 0.032). La presencia de varicocele se asoció con el bajo potencial fertilizante (OR= 5,27; IC 95 por ciento [1.34 - 24.09]). Conclusiones: El índice de fragmentación del ADN de la cromatina espermática afecta negativamente la calidad de los embriones a transferir. La edad y el varicocele se relacionan positivamente con el grado de fragmentación del ADN espermático. El consumo de alcohol, el hábito de fumar y la exposición a agentes físicos y químicos no se relacionaron con el índice de fragmentación del ADN espermático en este estudio(AU)


ABSTRACT Introduction: The factors that affect the spermatic DNA are being studied at present and in what extent they affect the implementation of high technology assisted reproduction techniques. In vitro fertilization is a topic of interest that has motivated many researchers. Objective: To determine the relationship between the rate of DNA fragmentation of sperm chromatin and the results of in vitro fertilization technique in infertile couples. Methods: A descriptive cross-sectional study was conducted in 107 infertile couples referred to the National Program of Attention to the Infertile Couple, where it was specified the association between the potential fertilizer and outcome variables: rate of fertilization, embryo quality and pregnancy. In addition, there were studied some factors that may influence the degree of DNA fragmentation. Results: The fertilizing potential was not related to the rate of fertilization, or with the clinical pregnancy, but it did with the embryo quality (p= 0.036). The paternal age proved to be statistically significant in relation to the fertilizing potential (p= 0.032). The presence of varicocele was associated with the low fertilizing potential (OR = 5.27; CI 95 percent [1.34 - 24.09]). Conclusions: The rate of DNA fragmentation of sperm chromatin negatively affects the quality of the embryos to be transferred. The age and varicocele are positively related with the degree of sperm DNA fragmentation. The consumption of alcohol, the smoking habit and the exposure to chemical and physical agents were not associated with the rate of sperm DNA fragmentation in this study(AU)


Subject(s)
Humans , Male , Female , Adult , Varicocele/etiology , Fertilization in Vitro/adverse effects , Reproductive Techniques/adverse effects , DNA Fragmentation , Epidemiology, Descriptive , Cross-Sectional Studies , Quality Management
6.
Article in English | WPRIM | ID: wpr-785645

ABSTRACT

OBJECTIVE: The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST).METHODS: Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern (“a”–“g”) were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed.RESULTS: The HOST examinations showed that >93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type “d”, 98.67% in type “g”, and 98.17% in type “f” spermatozoa.CONCLUSION: We found that the type “d” spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.


Subject(s)
Centrifugation, Density Gradient , Chromatin , DNA Fragmentation , DNA , Humans , Infertility , Semen , Semen Preservation , Sperm Head , Sperm Injections, Intracytoplasmic , Spermatozoa , Tail , Tissue Donors
7.
Blood Research ; : 253-261, 2019.
Article in English | WPRIM | ID: wpr-785541

ABSTRACT

BACKGROUND: Fluoranthene (FR) is a common environmental pollutant that exists in a complex mixture with other polycyclic aromatic hydrocarbons (PAHs). We identified biomarkers for monitoring FR exposure and investigated the rescue effect of FR-induced cellular toxicity via aryl hydrocarbon receptor (AHR) antagonist activity in bone marrow derived mesenchymal stem cells (BM-MSCs).METHODS: Morphological changes, viability, and rescue effects of an AHR antagonist (CH223191) were examined in BM-MSCs after exposure to FR. Cytotoxic effects were assayed using the tetrazolium-based colorimetric assay. Apoptosis was measured by annexin V and propidium iodide dye-based flowcytometry assay, mitochondrial membrane potential assay, and nuclear DNA fragmentation assay. Molecular signaling pathways of apoptosis and autophagy were investigated using immunoblotting. Proteomics were performed in order to reveal the spectra of cellular damage and identify biomarkers for FR exposure.RESULTS: Exposing BM-MSCs to FR (IC₅₀=50 µM) induced cell death and morphological changes, while the AHR antagonist showed rescue effects. Autophagy was activated and mitochondrial membrane potential was decreased. Proteomic analysis identified 48 deregulated proteins (26 upregulated and 22 downregulated). Among them, annexin A6, pyruvate kinase, UDP-glucose dehydrogenase, and phospholipase A2 could be potential biomarkers for FR exposure.CONCLUSION: The exposure of BM-MSCs to FR induced remarkable alterations in cellular biology and the proteome, allowing for identification of novel biomarkers for FR exposure. Furthermore, AHR antagonists might be able to prevent cellular damage due to FR exposure.


Subject(s)
Annexin A5 , Annexin A6 , Apoptosis , Autophagy , Biomarkers , Bone Marrow , Cell Death , DNA Fragmentation , Immunoblotting , Membrane Potential, Mitochondrial , Mesenchymal Stem Cells , Oxidoreductases , Phospholipases A2 , Polycyclic Aromatic Hydrocarbons , Propidium , Proteome , Proteomics , Pyruvate Kinase , Receptors, Aryl Hydrocarbon
8.
Yonsei Medical Journal ; : 461-466, 2019.
Article in English | WPRIM | ID: wpr-742559

ABSTRACT

PURPOSE: To investigate the associations between sperm DNA fragmentation (SDF) and embryo formation rate in normal responder women to in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). MATERIALS AND METHODS: Fifty-three consecutive, fresh IVF/ICSI cycles performed from 2014 to 2017 were selected. All women were normal responders (4 to 14 mature oocytes were retrieved) and at least one normally fertilized oocyte with two pronuclei was obtained in all cycles. Semen was collected on the day of oocyte retrieval, and SDF levels were measured by sperm chromatin dispersion test (Halosperm assay). At day 3 after insemination, embryo quality was evaluated by morphologic criteria and categorized as A/B/C/D. Top quality embryo were defined as grade A embryos with seven cells or more. RESULTS: SDF levels showed a positive linear correlation with the male's age (r=0.307, p=0.025) and a negative linear correlation with sperm motility (r=−0.491, p70%, the cut-off value SDF was <30.7% for each. Among individuals with SDF <30.7%, the median top-quality or grade A embryo formation rate was significantly higher than that among individuals with SDF ≥30.7% (38.1% vs. 20.0%, p=0.038; 50% vs. 25.0%, p=0.017). CONCLUSION: In normal responder women, high SDF level resulted in low day 3 embryo formation rates. Our results suggest a paternal effect on embryo quality in IVF/ICSI cycles.


Subject(s)
Chromatin , DNA Fragmentation , DNA , Embryonic Structures , Female , Fertilization in Vitro , Humans , In Vitro Techniques , Insemination , Oocyte Retrieval , Oocytes , Semen , Sperm Injections, Intracytoplasmic , Sperm Motility , Spermatozoa
9.
Article in English | WPRIM | ID: wpr-719643

ABSTRACT

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated IC50 value of 3 µM after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G1 phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the PERK/elF2α/CHOP apoptotic pathway, and mitochondrial Ca2+ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER-and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.


Subject(s)
Apoptosis , bcl-2-Associated X Protein , Caspase 9 , Cell Death , Cell Line , Colon , Colonic Neoplasms , DNA Fragmentation , Endoplasmic Reticulum , Extracellular Vesicles , Humans , Inhibitory Concentration 50 , Lymphoma, B-Cell , Mitochondria , Mitochondrial Membranes , Protein Kinases
10.
Article in Korean | WPRIM | ID: wpr-740552

ABSTRACT

PURPOSE: The aim of this study was to estimate the antioxidant activities of 50%, 70%, and 100% ethanol extracts of Lycium barbarum leaf and chlorophyll removal extract. METHODS: The antioxidant activities were estimated by measuring total polyphenol content and by assays of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfate) (ABTS) radical scavenging activities and ferric reducing antioxidant power (FRAP). In addition, reactive oxygen species (ROS) production, DNA fragmentation, and antioxidant enzyme (superoxide dismutase and catalase) activities of the extracts were measured in hydrogen peroxide (H2O2)-stressed HepG2 cells. RESULTS: The total polyphenol content, DPPH and ABTS radical scavenging activities, and FRAP value of the extracts increased in an ethanol concentration-dependent manner. The antioxidant activities of the chlorophyll-removal extracts were much higher than those of the chlorophyll-containing extracts. Cytotoxicity was not observed in HepG2 cells with extracts up to 1,000 µg/mL. All extracts inhibited ROS production in a concentration-dependent manner from 31.3 µg/mL and inhibited DNA damage at 250 µg/mL. The SOD and catalase activities of cell lines treated with the extracts and H2O2 were similar to those of normal cells, indicating a strong protective effect. CONCLUSION: Lycium barbarum leaf extracts had high antioxidant activities and protected H2O2-stressed HepG2 cells. Since the chlorophyll-removal extract exhibited higher antioxidant activities than the chlorophyll-containing ones and the cytoprotective effect was similar, chlorophyll removal extract of Lycium barbarum leaf could be developed as ingredients of functional food and cosmetics.


Subject(s)
Catalase , Cell Line , Chlorophyll , DNA Damage , DNA Fragmentation , Ethanol , Functional Food , Hep G2 Cells , Hydrogen Peroxide , Lycium , Reactive Oxygen Species
11.
Article in English | WPRIM | ID: wpr-742360

ABSTRACT

Male infertility (MI) is a complex multifactorial disease, and idiopathic infertility accounts for 30% of cases of MI. At present, the evidence for the effectiveness of empirical drugs is limited, and in vitro fertilization is costly and may increase the risk of birth defects and childhood cancers. Therefore, affected individuals may feel obliged to pursue natural remedies. Traditional Chinese medicine (TCM) may represent a useful option for infertile men. It has been demonstrated that TCM can regulate the hypothalamic-pituitary-testicular axis and boost the function of Sertoli cells and Leydig cells. TCM can also alleviate inflammation, prevent oxidative stress, reduce the DNA fragmentation index, and modulate the proliferation and apoptosis of germ cells. Furthermore, TCM can supply trace elements and vitamins, ameliorate the microcirculation of the testis, decrease the levels of serum anti-sperm antibody, and modify epigenetic markers. However, the evidence in favor of TCM is not compelling, which has hindered the development of TCM. This review attempts to elucidate the underlying therapeutic mechanisms of TCM. We also explore the advantages of TCM, differences between TCM and Western medicine, and problems in existing studies. Subsequently, we propose solutions to these problems and present perspectives for the future development of TCM.


Subject(s)
Apoptosis , Congenital Abnormalities , DNA Fragmentation , Epigenomics , Fertilization in Vitro , Germ Cells , Humans , Infertility , Infertility, Male , Inflammation , Leydig Cells , Male , Male , Medicine, Chinese Traditional , Microcirculation , Oxidative Stress , Sertoli Cells , Testis , Trace Elements , Vitamins
12.
Article in English | WPRIM | ID: wpr-764069

ABSTRACT

BACKGROUND AND OBJECTIVES: This study aims to explore the effects of a long non-coding RNA, LINC00525, on colorectal cancer (CRC) and its underlying molecular mechanisms. METHODS: The qPCR, MTT, colony formation, Western blotting, Luciferase reporter and biotin pull-down, shRNA knockdown and DNA fragmentation assays were performed in this study. RESULTS: High expressions of LINC00525 were associated with poor prognosis of CRC patients. LINC00525 knockdown decreased stemness properties and increased sensitivities to oxaliplatin. MiR-507 was a direct target of LINC00525 and overexpression of miR-507 significantly decreased abilities of tumorsphere formation and cell growth. Overexpression of miR-507 resulted in a decrease of expression of cancer stem cell markers and the increase of apoptosis rates. MiR-507 regulated the expression of ELK3. In addition, LINC00525 knockdown decreased the expression of ELK3. Restoration of ELK3 expression abrogated the effects of LINC00525 knockdown. LINC00525 could be served as prognostic marker of CRC. CONCLUSIONS: LINC00525 enhanced stemness properties and increased sensitivities of CRC cells to oxaliplatin by targeting miR-507/ELK3 axis.


Subject(s)
Apoptosis , Biotin , Blotting, Western , Colorectal Neoplasms , DNA Fragmentation , Humans , Luciferases , Neoplastic Stem Cells , Prognosis , RNA, Long Noncoding , RNA, Small Interfering
13.
Article in English | WPRIM | ID: wpr-763489

ABSTRACT

The objective of this study was to assess the association between dietary antioxidant intake and semen quality parameters in infertile men. In this cross-sectional study, dietary antioxidant intake was evaluated in 175 infertile Iranian men by a validated dish-based 106-item semi-quantitative food frequency questionnaire. Men were asked to abstain from ejaculation for at least 72 hours before sample collection. Semen parameters were assessed by a sperm counting chamber and Terminal deoxynucleotidyl transferase dUTP nick end labeling assay methods. Linear quantile regression was used to determine the associations between antioxidant nutrient intake and semen quality parameters (including total sperm count, sperm density, total motility, DNA damage and DNA fragmentation). Mean age of study participants was 32.19 ± 2.34 years. Compared with the lowest quartile, men in the highest quartile of dietary β-carotene and vitamin C intake had lower sperm DNA fragmentation index (Ptrend = 0.042 and Ptrend = 0.03, respectively). Also, dietary intake of beta-cryptoxanthin had a positive association with sperm density (Ptrend = 0.02), and dietary lutein was associated with total sperm count (P(trend) = 0.045). Dietary intake of other antioxidants did not significantly correlate with the indicators related to the quantity and quality of sperm (p > 0.05). These data suggest that dietary intake of some of the antioxidants is associated with semen related parameters.


Subject(s)
Antioxidants , Ascorbic Acid , Cross-Sectional Studies , Cryptoxanthins , DNA , DNA Damage , DNA Fragmentation , DNA Nucleotidylexotransferase , Ejaculation , Humans , Infertility , Lutein , Male , Oxidative Stress , Semen Analysis , Semen , Sperm Count , Spermatozoa
14.
Article in English | WPRIM | ID: wpr-763355

ABSTRACT

OBJECTIVE: Sperm cryopreservation has been widely used in assisted reproductive technology, as it offers great potential for the treatment of some types of male infertility. However, cryopreservation may result in changes in membrane lipid composition and acrosome status, as well as reductions in sperm motility and viability. This study aimed to evaluate sperm DNA fragmentation damage caused by conventional freezing using the sperm chromatin dispersion test. METHODS: In total, 120 fresh human semen samples were frozen by conventional methods, using SpermFreeze Solution as a cryoprotectant. Routine semen analysis and a Halosperm test (using the Halosperm kit) were performed on each sample before freezing and after thawing. Semen parameters and sperm DNA fragmentation were compared between these groups. RESULTS: There was a significant decrease in sperm progressive motility, viability, and normal morphology after conventional freezing (32.78%, 79.58%, and 3.87% vs. 16%, 55.99%, and 2.55%, respectively). The sperm head, midpiece, and tail defect rate increased slightly after freezing. Furthermore, the DNA fragmentation index (DFI) was significantly higher after thawing than before freezing (19.21% prior to freezing vs. 22.23% after thawing). Significant increases in the DFI after cryopreservation were observed in samples with both normal and abnormal motility and morphology, as well as in those with normal viability. CONCLUSION: Conventional freezing seems to damage some sperm parameters, in particular causing a reduction in sperm DNA integrity.


Subject(s)
Acrosome , Chromatin , Cryopreservation , DNA Fragmentation , DNA , Freezing , Humans , Infertility, Male , Male , Membranes , Reproductive Techniques, Assisted , Semen , Semen Analysis , Sperm Head , Sperm Motility , Spermatozoa , Tail
15.
Article in English | WPRIM | ID: wpr-763350

ABSTRACT

OBJECTIVE: Density gradient centrifugation (DGC) is frequently used to isolate high-motility fractions of spermatozoa. We compared the efficacy of four DGC media in terms of the percentage of morphologically normal spermatozoa, DNA fragmentation level, and hyaluronic acid (HA) binding ability. METHODS: Thirty men with a total motile spermatozoa count >80 million participated. Semen samples were divided into four aliquots, which were processed using PureSperm, PureCeption, Sidney, and SpermGrad media, respectively. The DNA fragmentation level was measured using the Halosperm assay kit and HA binding ability was measured using the HBA assay kit. RESULTS: The mean percentage of morphologically normal spermatozoa was significantly enhanced after DGC using all four media (10.3%, 9.9%, 9.8%, and 10.7%, respectively; p0.05). HA binding ability did not change after DGC using any of the four media. CONCLUSION: The four media were equally effective for obtaining a sperm fraction with highly motile, morphologically normal sperm. PureSperm, PureCeption, and SpermGrad media were equally effective for acquiring a sperm fraction with less DNA fragmentation.


Subject(s)
Centrifugation, Density Gradient , DNA Fragmentation , DNA , Humans , Hyaluronic Acid , Male , Semen , Spermatozoa
16.
Article in English | WPRIM | ID: wpr-763349

ABSTRACT

OBJECTIVE: The usual seminal profile has been customarily used for diagnosing male infertility based on an examination of semen samples. However, sperm DNA fragmentation has also been causally linked to reproductive failure, suggesting that it should be evaluated as part of male infertility assessments. To compare the ability of the five most widely utilized methodologies of measuring DNA fragmentation to predict male infertility and reactive oxygen species by Oxisperm kit assay. METHODS: In this case-control study, which received ethical committee approval, the participants were divided into fertile and infertile groups (50 patients in each group). RESULTS: The alkaline comet test showed the best ability to predict male infertility, followed by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay, the sperm chromatin dispersion (SCD) test, and the sperm chromatin structure assay (SCSA), while the neutral comet test had no predictive power. For our patient population, the projected cut-off point for the DNA fragmentation index was 22.08% using the TUNEL assay, 19.90% using SCSA, 24.74% using the SCD test, 48.47% using the alkaline comet test, and 36.37% using the neutral comet test. Significant correlations were found between the results of the SCD test and those obtained using SCSA and TUNEL (r =0.70 and r =0.68, respectively; p<0.001), and a statistically significant correlation was also found between the results of SCSA and the TUNEL assay (r =0.77, p<0.001). Likewise, the results of the alkaline comet test showed significant correlations with those of the SCD, SCSA, and TUNEL tests (r =0.59, r =0.57, and r =0.72, respectively; p<0.001). CONCLUSION: The TUNEL assay, SCSA, SCD, and the alkaline comet test were effective for distinguishing between fertile and infertile patients, and the alkaline comet test was the best predictor of male infertility.


Subject(s)
Case-Control Studies , Chromatin , DNA Fragmentation , DNA Nucleotidylexotransferase , DNA , Humans , In Situ Nick-End Labeling , Infertility , Infertility, Male , Male , Male , Methods , Reactive Oxygen Species , Semen , Sensitivity and Specificity , Spermatozoa
17.
Article in English | WPRIM | ID: wpr-763348

ABSTRACT

OBJECTIVE: As paternal age increases, the quality of sperm decreases due to increased DNA fragmentation and aneuploidy. Higher levels of structural chromosomal aberrations in the gametes ultimately decrease both the morphologic quality of embryos and the pregnancy rate. In this study, we investigated whether paternal age affected the euploidy rate. METHODS: This study was performed using the medical records of patients who underwent in vitro fertilization (IVF) procedures with preimplantation genetic screening (PGS) from January 2016 to August 2017 at a single center. Based on their morphological grade, embryos were categorized as good- or poor-quality blastocysts. The effects of paternal age were elucidated by adjusting for maternal age. RESULTS: Among the 571 total blastocysts, 219 euploid blastocysts were analyzed by PGS (38.4%). When the study population was divided into four groups according to both maternal and paternal age, significant differences were only noted between groups that differed by maternal age (group 1 vs. 3, p=0.031; group 2 vs. 4, p=0.027). Further analysis revealed no significant differences in the euploidy rate among the groups according to the morphological grade of the embryos. CONCLUSION: Paternal age did not have a significant impact on euploidy rates when PGS was performed. An additional study with a larger sample size is needed to clarify the effects of advanced paternal age on IVF outcomes.


Subject(s)
Aneuploidy , Blastocyst , Chromosome Aberrations , DNA Fragmentation , Embryonic Development , Embryonic Structures , Female , Fertilization in Vitro , Genetic Testing , Germ Cells , Humans , In Vitro Techniques , Maternal Age , Medical Records , Paternal Age , Pregnancy , Pregnancy Rate , Sample Size , Spermatozoa
18.
Yonsei Medical Journal ; : 509-516, 2019.
Article in English | WPRIM | ID: wpr-762085

ABSTRACT

PURPOSE: This study was conducted to verify the induction and mechanism of selective apoptosis in G361 melanoma cells using anti-HER2 antibody-conjugated gold nanoparticles (GNP-HER2). MATERIALS AND METHODS: Following GNP-HER2 treatment of G361 cells, cell cycle arrest and apoptosis were measured by WST-1 assay, Hemacolor staining, Hoechst staining, immunofluorescence staining, fluorescence-activated cell sorting analysis, and Western blotting.


Subject(s)
Actins , Apoptosis Inducing Factor , Apoptosis , Blotting, Western , Caspase 3 , Caspases , Cell Adhesion , Cell Cycle , Cell Cycle Checkpoints , Cell Death , Cyclin A , Cyclin D1 , Cyclin E , Cyclins , Cytochromes c , Cytoplasm , DNA Fragmentation , Down-Regulation , Flow Cytometry , Fluorescent Antibody Technique , Focal Adhesions , Melanoma , Mitochondria , Nanoparticles , Phosphotransferases , ErbB Receptors , Up-Regulation
19.
Article in English | WPRIM | ID: wpr-760676

ABSTRACT

OBJECTIVE: The male reproductive system generates, accumulates, and transports the sperm. In this study, 2 methods of surgically retrieving sperm, namely, testicular sperm aspiration (TESA) and percutaneous epididymal sperm aspiration (PESA), are discussed and studied in men aged ≤38 years to achieve successful conception using assisted reproductive technology. The purpose was to assess the fertilization rate (FA), clinical pregnancy, and live birth rate (LBR) with sperm. METHODS: A total of 287 semen samples were divided into 4 groups as follows: fresh PESA (n=73), frozen PESA (n=65), fresh TESA (n=128), and frozen TESA (n=21). The DNA fragmentation test using sperm chromatin dispersion assay was measured and reported. RESULTS: FA was 70.3% and 65.5%, (P<0.022) for fresh and frozen epididymal sperm and 53.8% and 49.5%, (P<0.032) for fresh and frozen testicular sperm. LBR was 33.6% and 30.2% (P<0.075) for fresh and frozen epididymal sperm (PESA) and 22.7% and 18.2% (P<0.063) for fresh and frozen-thawed TESA sperm. CONCLUSION: Exposure to tissue shearing may adversely affect sperm quality. Increased sperm DNA damage due to long-term exposure while teasing enhances reactive oxygen species production foremost to membrane damage because of the oxidation of polyunsaturated fatty acid in lipids (lipid peroxidation), oxidation of amino acid in proteins, and inactivation of specific enzymes, all leading to enzymatic dipping and possibility of less fertilization and conception as indicated by the increase in LBR with fresh/frozen PESA compared to with fresh/frozen TESA.


Subject(s)
Chromatin , DNA Damage , DNA Fragmentation , Fertilization , Humans , Infertility , Live Birth , Male , Membranes , Pregnancy , Reactive Oxygen Species , Reproductive Techniques, Assisted , Retrospective Studies , Semen , Sperm Injections, Intracytoplasmic , Sperm Retrieval , Spermatozoa
20.
Article in English | WPRIM | ID: wpr-785835

ABSTRACT

BACKGROUND AND OBJECTIVES: Oxidative stress (OS) is known to be an important factor of male infertility. Adipose-derived mesenchymal stem cells (AD-MSCs) are known to have immune-modulatory and anti-oxidant effects through their secretions, hence raising the idea of their potential benefit to improve sperm parameters. This study aims at investigating the effect of AD-MSCs conditioned medium (CM) on human sperm parameters in the presence and absence of H2O2-induced OS.METHODS AND RESULTS: Sperm samples were collected from 30 healthy men and divided into two groups: non-stressed and H2O2-stressed. Isolated AD-MSCs from healthy donors undergoing liposuction were cultured and CM was collected at 24, 48 and 72 h. Both sperm groups were cultured with CM and a time course was performed followed by an evaluation of sperm parameters. The incubation of non-stressed and stressed sperm samples with AD-MSCs-CM for 24 h was found to have the optimum impact on sperm vacuolization, DNA fragmentation and OS levels in comparison to other incubation timings, while preserving motility, viability and morphology of cells. Incubation with CM improved all sperm parameters except morphology in comparison to the non-treated group, with the best effect noted with CM collected at 24 h rather than 48 or 72 h for sperm vacuolization and DNA fragmentation. When compared to fresh semen parameters (T0), samples cultured with CM 24 h showed a significant decrease in sperm vacuolization and DNA fragmentation while keeping other parameters stable.CONCLUSIONS: AD-MSCSs-CM improves sperm quality, and hence can be used in treating infertility and subsequently enhancing IVF outcomes.


Subject(s)
Antioxidants , Culture Media, Conditioned , DNA Fragmentation , DNA , Humans , In Vitro Techniques , Infertility , Infertility, Male , Lipectomy , Male , Mesenchymal Stem Cells , Oxidative Stress , Semen , Spermatozoa , Tissue Donors
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