Publications by authors named "De Yi Liu"

28 Publications

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Commentary on "Altered and mRNA expression in ejaculated spermatozoa of men with impaired sperm characteristics".

Authors:
De-Yi Liu

Asian J Androl 2018 May-Jun;20(3):318

Andrology Department, Melbourne IVF and Department of Obstetrics and Gynecology, Royal Women's Hospital, The University of Melbourne, Melbourne, Victoria 3002, Australia.

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http://dx.doi.org/10.4103/aja.aja_67_17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5952493PMC
January 2019

[Blastocystis Infection in Pet Dogs in Selected Areas of Anhui and Zhejiang Provinces].

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 2016 Jun;34(3):272-6

Objective: To determine the prevalence and subtypes of Blastocystis in pet dogs in Anhui and Zhejiang Provinces.

Methods: Pet dog fecal samples were collected from veterinary clinics in Baohe District of Hefei City, Xuanzhou District of Xuancheng City, Fengyang County and Mingguang City of Chuzhou City, Longzihu District of Bengbu City, Si County of Suzhou City in Anhui Province, as well as in Yuhang District of Hangzhou City in Zhejiang Province from April to December, 2013. Blastocystis was detected by microscopic examination as well as nested PCR and single-step PCR based on small subunit ribosomal DNA sequences. The PCR products were sequenced, and Blastocystis subtypes were assigned.

Results: A total of 315 pet dog fecal samples were collected. The incidence of Blastocystis infection determined by microscopic examination, nested PCR and single-step PCR was 1.3%(4/315), 1.9%(6/315) and 1.9%(6/315), respectively. In addition, the two PCR methods both revealed Blastocystis infection only in Chuzhou City(3.4%, 1/29) and Hefei City(5.6%, 4/72). The pet dogs in Chuzhou City carried ST1 and ST2 subtypes while those in Hefei carried only ST1. There was no significant difference in infection rate between adult and young dogs and between male and female dogs.

Conclusion: Blastocystis infection exists in pet dogs in Chuzhou City and Hefei City.
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June 2016

[Molecular Detection of Giardia lamblia and Cryptosporidium Species in Pet Dogs].

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 2015 Oct;33(5):362-7

Objective: To determine the prevalence of Giardia lamblia and Cryptosporidium species infection in pet dogs, and identify the G. lamblia assemblages and Cryptosporidium species.

Methods: A total of 315 fresh fecal samples were collected from pet clinics in five counties of Anhui Province and in Hangzhou of Zhejiang Province. Hemi-nested-PCR targeting the GDH gene of G. lamblia and nested-PCR targeting the SSU rRNA gene of Cryptosporidium were performed in all the fecal samples. The PCR products were sequenced and analyzed using bioinformatics methods to identify the G. lamblia assemblages and Cryptosporidium species.

Results: The positive rates of G. lamblia and Cryptosporidium spp. infections in the 315 fecal samples were 3.2% (10/315) and 1.6% (5/315), respectively. Specifically, the two indicators were both significantly higher in dogs ≤12 months (17.8% and 11.1%, respectively) than in adult dogs (0.7% and 0.0%)(P<0.05). However, there was no significant difference in the two indicators between male and female dogs. In addition, two G. lamblia assemblages were identified, assemblages B (n=6) and D (n=4). Sequence analysis of PCR products of the SSU rRNA gene showed that the five Cryptosporidium isolates were C. canis (n =5).

Conclusion: The prevalences of G. lamblia and Cryptosporidium infection in pet dogs in Anhui and Zhejiang Provinces were 3.2 % and 1.6 %, respectively. The assemblages of G. lamblia in this study are of types B and D.
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October 2015

[Prevalence of Neospora caninum Infection in the Intestine of Pet Dogs in Some Areas of Anhui and Zhejiang].

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 2015 Aug;33(4):283-6

Objective: To investigate the prevalence of Neospora caninum infection in the intestine of pet dogs in areas of Anhui and Zhejiang.

Methods: A total of 315 fecal samples from pet dogs were collected in pet clinics from April to December 2013 in Baohe District in Hefei city, Xuanzhou District in Xuancheng city, Fengyang County in Chuzhou city, Longzihu District in Bengbu city, and Si County in Suzhou City in Anhui Province, as well as in Yuhang District in Hangzhou city of Zhejiang province. All samples underwent nested-PCR targeting Neospora-specific gene NCLI-004830. The results were further confirmed by PCR amplification of N. caninum ITS1 followed by sequence analysis.

Results: The rate of N. caninum infection in the 315 samples was 1.59% (5/315). The infection rate in Chuzhou and Bengbu was 3.37% and 6.45%, respectively, and no N. caninum infection was found in the remaining areas. There was no association between the infection rate and the sex or age of the dogs.

Conclusion: N. caninum infection is prevalent in pet dogs in Chuzhou and Bengbu of Anhui.
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August 2015

The Physiology of BDNF and Its Relationship with ADHD.

Mol Neurobiol 2015 Dec 30;52(3):1467-1476. Epub 2014 Oct 30.

Key Laboratory of Environment and Health, Ministry of Education & Department of Epidemiology and Biostatistics, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People's Republic of China.

Brain-derived neurotrophic factor (BDNF) is a major neurotrophin in the central nervous system that plays a critical role in the physiological brain functions via its two independent receptors: tropomyosin-related kinase B (TrkB) and p75, especially in the neurodevelopment. Disrupting of BDNF and its downstream signals has been found in many neuropsychological diseases, including attention-deficit hyperactivity disorder (ADHD), a common mental disorder which is prevalent in childhood. Understanding the physiological functions of BDNF during neural development and its potential relationship with ADHD will help us to elucidate the possible mechanisms of ADHD and to develop therapeutic approaches for this disease. In this review, we summarized the important literatures for the physiological functions of BDNF in the neurodevelopment. We also performed an association study on the functional genetic variation of BDNF and ADHD by a case-control study in the Chinese mainland population and revealed the potential correlation between BDNF and ADHD which needs further research to confirm.
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http://dx.doi.org/10.1007/s12035-014-8956-6DOI Listing
December 2015

Daidzein enhances immune function in late lactation cows under heat stress.

Anim Sci J 2014 Jan 17;85(1):85-9. Epub 2013 Jun 17.

Anhui Science and Technology University, Bengbu, China.

Heat stress decreases natural immunity making cows more vulnerable to diseases. A previous study reported that daidzein can enhance animal resistance to heat stress and regulate animal immunocompetence. However, it is unclear whether daidzein regulates the immune performance of late lactation cows under heat stress. In this study, late lactation cows in four groups were raised in hot weather and fed with basic diet, basic diet plus 200, 300, 400 mg/day daidzein, respectively, and the experimental period was 60 days. Blood was collected to examine the changes of serum total protein (TP), albumin (ALB), immunoglobulin G (IgG), interferon alpha (IFN-α), and interleukin-2 (IL-2). We found the levels of serum IgG and INF-α were significantly higher in late lactation cows after 300 and 400 mg/day daidzein treatment compared to those in the control group and 200 mg/day daidzein treatment (P < 0.05 or P < 0.01). Moreover, 300 and 400 mg/day daidzein treatment markedly increased serum IL-2 (P < 0.01), while the levels of serum TP and ALB were not changed by any concentration of daidzein treatment (P > 0.05). Daidzein can enhance the immunocompetence of late lactation cows and strengthen cow resistance to heat stress.
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http://dx.doi.org/10.1111/asj.12080DOI Listing
January 2014

The molecular chaperone HSPA2 plays a key role in regulating the expression of sperm surface receptors that mediate sperm-egg recognition.

PLoS One 2012 29;7(11):e50851. Epub 2012 Nov 29.

Priority Research Centre in Reproductive Science, School of Environmental and Life Sciences, Discipline of Biological Sciences, The University of Newcastle, Callaghan, New South Wales, Australia.

A common defect encountered in the spermatozoa of male infertility patients is an idiopathic failure of sperm-egg recognition. In order to resolve the molecular basis of this condition we have compared the proteomic profiles of spermatozoa exhibiting an impaired capacity for sperm-egg recognition with normal cells using label free mass spectrometry (MS)-based quantification. This analysis indicated that impaired sperm-zona binding was associated with reduced expression of the molecular chaperone, heat shock 70 kDa protein 2 (HSPA2), from the sperm proteome. Western blot analysis confirmed this observation in independent patients and demonstrated that the defect did not extend to other members of the HSP70 family. HSPA2 was present in the acrosomal domain of human spermatozoa as a major component of 5 large molecular mass complexes, the most dominant of which was found to contain HSPA2 in close association with just two other proteins, sperm adhesion molecule 1 (SPAM1) and arylsulfatase A (ARSA), both of which that have previously been implicated in sperm-egg interaction. The interaction between SPAM1, ARSA and HSPA2 in a multimeric complex mediating sperm-egg interaction, coupled with the complete failure of this process when HSPA2 is depleted in infertile patients, provides new insights into the mechanisms by which sperm function is impaired in cases of male infertility.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0050851PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3510172PMC
May 2013

Clinical value of sperm DNA damage should be assessed in motile sperm fraction rather than whole ejaculated sperm.

Fertil Steril 2013 Feb 22;99(2):367-71. Epub 2012 Oct 22.

Melbourne IVF, Melbourne, Victoria, Australia.

Objective: To determine differences and frequency of excessive (≥50%) sperm DNA damage between ejaculated and motile sperm.

Design: Sperm DNA damage was assessed by acridine orange fluorescence and the results of ejaculated and motile sperm were compared.

Setting: Public and private clinical assisted reproduction centers.

Patient(s): A total of 272 subfertile men were studied.

Intervention(s): None.

Main Outcome Measure(s): Semen analysis and sperm DNA damage.

Result(s): Sperm DNA damage was negatively correlated with sperm motility and normal morphology. Overall, 39.7% (108 of 272) of semen samples had excessive sperm DNA damage. In contrast, only 15% (41 of 272) of motile sperm fractions had excessive DNA damage. Based on DNA results of motile sperm and semen characteristics, the proportion of men with excessive sperm DNA damage was 26% in severe teratozoospermia, 17.5% in oligozoospermia, 12.5% in moderate teratozoospermia, and 4.6% in normozoospermia. Severe teratozoospermia had five times more frequent excessive DNA damage than normozoospermia.

Conclusion(s): Abnormal sperm morphology is highly associated with sperm DNA damage. Results of DNA damage of ejaculated sperm do not accurately reflect DNA status of motile sperm. Therefore, sperm DNA damage should be assessed in motile sperm fraction rather than whole ejaculated sperm.
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http://dx.doi.org/10.1016/j.fertnstert.2012.10.005DOI Listing
February 2013

Defective protein kinase A and C pathways are common causes of disordered zona pellucida (ZP)--induced acrosome reaction in normozoospermic infertile men with normal sperm-ZP binding.

Fertil Steril 2013 Jan 15;99(1):86-91. Epub 2012 Sep 15.

Melbourne IVF, East Melbourne, Victoria, Australia; Department of Obstetrics and Gynaecology, Royal Women's Hospital, University of Melbourne, Parkville, Victoria, Australia.

Objective: To determine association between defective protein kinases C (PKC) and A (PKA) and disordered zona pellucida (ZP)-induced acrosome reaction (DZPIAR) in normozoospermic infertile men with normal sperm-ZP binding.

Design: Sperm from DZPIAR infertile men were treated without (control) or with (test) phorbol myristate acetate (PMA, PKC activator) or dibutyryl cyclic AMP (dbcAMP, PKA activator) under in vitro standard culture condition. The ZP-induced AR was assessed and compared between control and test.

Setting: Public and private hospital-based clinical assisted reproduction technology (ART) centers.

Patient(s): A total of 51 DZPIAR infertile men were involved in this study.

Intervention(s): None.

Main Outcomes Measure(s): Sperm-ZP binding and the ZP-induced IAR.

Result(s): Both PMA and dbcAMP enhanced ZP-induced AR up to a normal level (≥25%) in some subjects with DZPIAR: 29 (57%) with PMA and 27 (53%) with dbcAMP. Overall 35 (69%) had the ZP-induced AR enhanced to normal by PMA or dbcAMP but 16 (31%) had little or no response to either agent. Fourteen men responded to the two activators differently: 8 effective only with PMA and 6 effective only with dbcAMP.

Conclusion(s): Defective upstream of PKC and PKA pathways are highly associated with disordered ZPIAR in normozoospermic infertile men with normal sperm-ZP binding.
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http://dx.doi.org/10.1016/j.fertnstert.2012.08.040DOI Listing
January 2013

Quinn's advantage fertilization medium enhances zona pellucida-induced acrosome reaction compared with human tubal fluid medium.

Reprod Biomed Online 2011 Dec 27;23(6):735-9. Epub 2011 Jul 27.

Melbourne IVF, East Melbourne, Victoria 3002, Australia.

The aim of this study was to compare sperm velocity, hyperactivation, zona pellucida (ZP) binding and ZP-induced acrosome reaction (AR) between Quinn's advantage fertilization (QAF), human tubal fluid (HTF) and Ham's F10 media. Semen samples were obtained from normozoospermic men and motile spermatozoa were prepared by gradient centrifugation (PureSperm). Unfertilized oocytes from clinical IVF were used for spermatozoa-oocyte interaction tests. Sperm velocity and hyperactivation were assessed using a Hamilton-Thorn motility analyser. When media were supplemented with human albumin, sperm motility and velocity and sperm binding were not significantly different between QAF and HTF. However, ZP-induced AR was significantly higher with QAF than HTF (42±22 versus 21±18, P<0.th001). Sperm velocity, hyperactivation and sperm binding were also significantly higher in QAF than Ham's F10 media. Supplementation of media with either human serum or human albumin showed no difference in effect on all sperm test results. In conclusion, QAF medium significantly enhances ZP-induced AR which is essential for sperm penetration. Thus QAF appears to be a better medium than HTF for sperm fertilizing ability in conventional IVF.
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http://dx.doi.org/10.1016/j.rbmo.2011.07.013DOI Listing
December 2011

[Importance of early diagnosis and operative treatment for trans-scaphoid perilunate dislocation].

Zhongguo Gu Shang 2011 Feb;24(2):163-5

Department of Orthopaedics, the 309th Hospital of PLA, Beijing 100091, China.

Objective: To evaluate the effectiveness of open reduction and internal fixation and repair of palmar ligment in treating trans-scaphoid perilunate dislocation.

Methods: From June 1995 to June 2001,14 patients with trans-scaphoid perilunate dislocation were treated with open reduction and internal fixation and repair of palmar ligment. Among them,there were 13 males and 1 female,the ranging in age from 21 to 38 years,averaged 25.4 years. All patients were posterior dislocation and all operations were performed within 2 weeks after injury.

Results: All patients were followed up from 24 to 60 months with an average of 28.3 months. Thirteen scaphoid fractures were primary healed and functions of wrist joint were good. Bone disunion was found in 1 case and part functions of wrist joint were limited. No found necrosis of lunate and scaphoid. According to clinical scoring system of Cooney, 9 case got excellent results, 3 good, 1 fair and 1 poor.

Conclusion: Open reduction and internal fixation and repair of palmar ligament is effective in treating trans-scaphoid perilunate dislocation,which can early provide steady fixation for scaphoid,and profit to recover blood supply of lunatum and subterminal scaphoid.
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February 2011

Could using the zona pellucida bound sperm for intracytoplasmic sperm injection (ICSI) enhance the outcome of ICSI?

Authors:
De-Yi Liu

Asian J Androl 2011 Mar 17;13(2):197-8. Epub 2011 Jan 17.

Melbourne IVF, East Melbourne, Vic. 3002, Australia.

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http://dx.doi.org/10.1038/aja.2010.179DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3739213PMC
March 2011

Use of zona pellucida-bound sperm for intracytoplasmic sperm injection produces higher embryo quality and implantation than conventional intracytoplasmic sperm injection.

Fertil Steril 2011 Feb 25;95(2):815-8. Epub 2010 Oct 25.

Reproductive Medical Centre, Third Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi, People's Republic of China.

This goal of this study was to compare the outcomes of conventional intracytoplasmic sperm injection (ICSI; control group, n = 53 couples) and a modified ICSI technique using zona pellucida (ZP)-bound sperm for injection of oocytes (test group, n = 53 couples). The proportion of high-quality embryos (grades 1 and 2) and implantation rate were significantly higher in the test group than in the control group, but the difference in fetal heart pregnancy rate was not significant despite seven more pregnancies being obtained in the test group (26 pregnancies) versus the control group (19 pregnancies) following fresh embryo transfers.
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http://dx.doi.org/10.1016/j.fertnstert.2010.09.015DOI Listing
February 2011

Comparison of outcomes of conventional intracytoplasmic sperm injection and intracytoplasmic sperm injection using sperm bound to the zona pellucida of immature oocytes.

Fertil Steril 2010 Feb 29;93(2):672-4. Epub 2009 Oct 29.

Reproductive Services and Melbourne IVF, Department of Obstetrics and Gynaecology, University of Melbourne, Royal Women's Hospital, Parkville, Victoria 3052, Australia.

This study was to compare the outcomes of conventional (scientist-selected sperm) intracytoplasmic sperm injection (ICSI) and a modified ICSI using zona pellucida (ZP)-bound sperm. Although with 39 couples in each group there was no statistical significance in fertilization, embryo development, implantation, and fetal heart clinical pregnancy rates, the higher implantation and clinical pregnancy rates with ZP-bound sperm encourages further evaluation with larger numbers of subjects using sibling oocytes to determine if the ZP-bound sperm are biologically and functionally superior.
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http://dx.doi.org/10.1016/j.fertnstert.2009.08.063DOI Listing
February 2010

Anti-human proacrosin antibody inhibits the zona pellucida (ZP)-induced acrosome reaction of ZP-bound spermatozoa.

Fertil Steril 2010 May 21;93(7):2456-9. Epub 2009 Oct 21.

Instituto de Biología y Medicina Experimental (IBYME), National Research Council of Argentina (CONICET), 1428ADN, Buenos Aires, Argentina.

The anti-acrosin monoclonal antibody AcrC5F10 inhibited proacrosin activation, proacrosin-human zona pellucida glycoprotein A (ZPA) binding, and the zona pellucida (ZP)-induced acrosome reaction of the ZP-bound spermatozoa but had no significant effect on sperm-ZP binding. These results suggest that proacrosin-acrosin may play an important role in the ZP-induced acrosome reaction of spermatozoa after primary binding to the ZP.
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http://dx.doi.org/10.1016/j.fertnstert.2009.08.040DOI Listing
May 2010

Relationship between seminal plasma zinc concentration and spermatozoa-zona pellucida binding and the ZP-induced acrosome reaction in subfertile men.

Asian J Androl 2009 Jul 11;11(4):499-507. Epub 2009 May 11.

Department of Obstetrics and Gynaecology, University of Melbourne, Royal Women's Hospital, Parkville 3052, Australia.

The aim of this study was to determine the relationship between seminal zinc concentration and spermatozoa-zona pellucida (ZP) binding and the ZP-induced acrosome reaction (ZPIAR) in subfertile men. Semen analyses and seminal zinc concentration assessments were carried out according to the World Health Organization manual for 458 subfertile men. A spermatozoa-ZP interaction test was carried out by incubating 2 x 10(6) motile spermatozoa with a group of four unfertilized oocytes obtained from a clinical in vitro fertilization programme. After 2 h of incubation, the number of spermatozoa bound per ZP and the ZPIAR of ZP-bound spermatozoa were examined. The effect of adding 0.5 mmol L(-1) zinc to the media on the ZPIAR of spermatozoa from normozoospermic men was also tested in vitro. Seminal zinc concentration positively correlated with sperm count and duration of abstinence, but negatively correlated with semen volume. On analysis of data from all participants, both spermatozoa-ZP binding and the ZPIAR were significantly correlated with sperm motility and normal morphology, but not with seminal zinc concentration. However, in men with normozoospermic semen, the seminal zinc concentration was significantly higher in men with defective ZPIAR (< 16%) than in those with normal ZPIAR (>or= 16%) (P < 0.01). The addition of 0.5 mmol L(-1) zinc to the culture media had no effect on spermatozoa-ZP binding, but significantly reduced the ZPIAR in vitro (P < 0.001). In conclusion, seminal zinc concentration is correlated with sperm count and the duration of abstinence in subfertile men. In men with normozoospermic semen, high seminal zinc concentration may have an adverse effect on the ZPIAR.
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http://dx.doi.org/10.1038/aja.2009.23DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3735308PMC
July 2009

Comparison of the frequency of defective sperm-zona pellucida (ZP) binding and the ZP-induced acrosome reaction between subfertile men with normal and abnormal semen.

Hum Reprod 2007 Jul 23;22(7):1878-84. Epub 2007 Apr 23.

Department of Obstetrics and Gynaecology, University of Melbourne, Royal Women's Hospital, 132, Grattan Street, Carlton, Victoria 3053, Australia.

Background: The aim of this study was to compare the frequency of defective sperm-zona pellucida (ZP) binding (DSZPB) and defective ZP-induced acrosome reaction (DZPIAR) in subfertile men (i.e. male partners of infertile couples) with normal and abnormal semen analyses.

Methods: A total of 1030 subfertile men with normal semen analysis (n=255), oligozoospermia (count<20x10(6)/ml, n=136), severe teratozoospermia (strict normal morphology
Results: For the groups of men with normal semen or mild-moderate teratozoospermia, severe teratozoospermia and oligozoospermia, the frequencies of DSZPB were: 13, 21, 29 and 28%, respectively, and in those normal SZPB, DZPIAR were 27, 36, 56 and 68%, respectively. Overall DSZPB and ZPIAR were 36, 49, 68 and 77% for the four groups, respectively. The highest frequencies of defective sperm-ZP interaction were in the oligozoospermia and severe teratozoospermia groups. In the normal and teratozoospermia groups, subjects with a relatively low sperm concentration (20-60x10(6)/ml) had a significantly higher frequency of DZPIAR.

Conclusion: Defective sperm-ZP interaction is a major mechanism of male infertility. DZPIAR is more frequent than DSZPB in subfertile men with either normal or abnormal semen, suggesting that sequential sperm-ZP interaction tests are essential to detect these sperm defects.
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http://dx.doi.org/10.1093/humrep/dem087DOI Listing
July 2007

[Assessment of human sperm function and clinical management of male infertility].

Zhonghua Nan Ke Xue 2007 Feb;13(2):99-109

Department of Obstetrics and Gynaecology, University of Melbourne, Melbourne IVF and Reproductive Services, Royal Womens Hospital, Melbourne, VIC 3053, Australia.

In this article, we provide an update review on the implication of the assessment of human sperm function and the management of male infertility in clinical assisted reproductive technology (ART) known as in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In most ART clinics, the assessment of male fertility is still mainly based on routine semen analysis but it is inaccurate in predicting sperm fertilizing ability. Thus it is often difficult to determine if IVF or ICSI will be an optimal treatment for patients in the initial cycle. Before introduction of ICSI, frequency of low ( <30%) fertilization rate in IVF was very high (20-35% of patients). Evidence suggests that sperm defects are the major contributors to complete failure of fertilization in IVF. Most common sperm defects are oligozoospermia, asthenozoospermia and teratozoospermia though many of the patients are shown to be normal in routine semen analysis. In the literature, many new sperm function tests have been developed, including sperm DNA normalities assessed by Acridine Orange (AO), sperm-zona pellucida (ZP) binding, the ZP-induced acrosome reaction (AR) , sperm-ZP penetration and recently hyaluronan binding assay (HBA). For routine semen analysis, sperm morphology is one of the most useful values for the prediction of sperm function but is also the most difficult test to perform accurately and consistently. Oocytes that failed to fertilize in clinical IVF/ICSI are valuable biological materials for testing sperm function. The human ZP selectively binds sperm with normal morphology and an intact acrosome. The ZP-induced AR is highly correlated with sperm-ZP penetration and disordered ZP-induced AR causes infertility in about 25% men with unexplained infertility with normal semen analysis. Both oligozoospermic (sperm count < 20 x 10(6) /ml) and severe teratozoospermia (strict normal sperm morphology < or =5%) men have a very high ( >70%) frequency of defective sperm-ZP interaction. Thus patients with defects of sperm-ZP interaction should be identified and treated with ICSI since they have high risk of low or zero fertilization rate in IVF. HBA test highly correlates with sperm motility and normal morphology but provides no additional information about sperm fertility. Clinical value of sperm DNA normalities detected by AO for the prediction of ART outcomes is currently still inconclusive and requires further investigation. In conclusion, addition of some of these new sperm tests to routine semen analysis could significantly improve the management of male infertility in clinical ART.
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February 2007

Recovery of human sperm motility and ability to interact with the human zona pellucida after more than 28 years of storage in liquid nitrogen.

Fertil Steril 2006 Sep 30;86(3):721-2. Epub 2006 Jun 30.

Andrology Unit, The Royal Women's Hospital, Melbourne, Victoria, Australia.

Six donor semen samples were evaluated after 28 years cryopreservation in liquid nitrogen. The results showed that the samples retained good postthaw motility recovery and normal levels of binding to the human zona pellucida and that four of the five samples tested also gave normal levels of zona-induced acrosome reaction. In conclusion, human sperm can survive very long-term storage, which is pertinent information for clinicians referring boys and young men for sperm banking before chemotherapy.
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http://dx.doi.org/10.1016/j.fertnstert.2006.01.050DOI Listing
September 2006

Relationship between human sperm-hyaluronan binding assay and fertilization rate in conventional in vitro fertilization.

Hum Reprod 2006 Jun 4;21(6):1545-50. Epub 2006 Apr 4.

Reproductive and Genetics Institute, Chongqing Obstetrics and Gynecology Hospital, Chongqing, China.

Background: Sperm-hyaluronan-binding assay (HBA) is one of the commercial kits being marketed for routine testing of sperm maturity and fertility. However, there is no report of whether the HBA can provide additional information over standard semen analysis for sperm-fertilizing ability. The objective of this study was to investigate the relationship between HBA and fertilization rate in conventional IVF.

Methods: A total of 175 IVF patients with > or = 3 mature oocytes inseminated were included in the study. Both the standard semen analysis and the HBA were performed on the same ejaculated sperm samples used for IVF treatments. Relationships between the semen analysis and the HBA results and fertilization rate were analysed by both the Spearman test and the multivariate logistic regression analysis.

Results: Both total and progressive sperm motility and normal morphology were highly correlated with HBA scores. While both normal sperm morphology and HBA scores were statistically significantly related to fertilization rates, the HBA was less significant than normal sperm morphology. The HBA does not provide additional information for identifying patients with a poor fertilization rate.

Conclusion: HBA is highly significantly correlated with sperm motility and morphology but is less significant than sperm morphology in relation to the fertilization rate in IVF. Thus, the clinical predictive value of HBA for sperm-fertilizing ability in vitro is limited.
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http://dx.doi.org/10.1093/humrep/del008DOI Listing
June 2006

Clinical application of sperm-oocyte interaction tests in in vitro fertilization--embryo transfer and intracytoplasmic sperm injection programs.

Fertil Steril 2004 Nov;82(5):1251-63

Department of Obstetrics and Gynecology and Reproductive Services, The Royal Women's Hospital, University of Melbourne, Melbourne, Victoria, Australia.

Objective: To review the clinical value of sperm-oocyte interaction tests for the diagnosis and management of infertility by standard IVF or intracytoplasmic sperm injection (ICSI).

Design: Review of recent publications on relationships among sperm-oocyte interaction tests, sperm characteristics, and results of IVF and determination of frequency of defective sperm-oocyte interaction in infertile men.

Main Outcome Measure(s): Fertilization rates with IVF, sperm characteristics, sperm-zona pellucida (ZP) binding, ZP-induced acrosome reaction (AR), and sperm-ZP penetration.

Result(s): Sperm defects associated with low sperm-ZP binding or impaired ZP-induced AR and sperm-ZP penetration are the major causes of failure of fertilization when all or most oocytes from a couple do not fertilize in standard IVF. There is a high frequency of defective sperm-ZP interaction in men with oligozoospermia (<20 x 10(6)/mL) and severe teratozoospermia (strict normal sperm morphology < or =5%). Sperm morphology correlates with sperm-ZP binding, and sperm concentration correlates with ZP-induced AR in infertile men with sperm concentrations >20 x 10(6)/mL. Defective ZP-induced AR may cause infertility in up to 25% men with idiopathic infertility. These patients require ICSI despite the normal standard semen analyses.

Conclusion(s): Sperm-oocyte interaction tests are useful for diagnosis of subtle sperm defects that cause infertility in men without severe abnormalities of semen analysis. Pre-IVF diagnosis of these sperm defects will assist in the clinical assignment of patients to treatment with either standard IVF or ICSI.
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http://dx.doi.org/10.1016/j.fertnstert.2003.10.057DOI Listing
November 2004

Flow cytometry and microscopic acridine orange test: relationship with standard semen analysis.

Reprod Biomed Online 2004 Apr;8(4):398-407

Clinical Sciences Centre, Medical Research Council, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK.

Improved prediction of male fertility requires advances in semen analysis. This study examined the reproducibility and independence of the flow cytometry acridine orange test (FCM-AOT) of sperm chromatin integrity as an assessment of semen quality. The study found that FCM-AOT results are not significantly affected by up to 6 h delay in semen preparation (n = 9) or contamination of semen with moderate concentrations of bacteria (<10(8)/ml E. coli or Staph. epidermidis, n = 14). The variation of replicate measurements within samples was low (%Abnormal alpha(t): SD = 1.4, 95%CI = 4.6, n = 25) and different samples from the same men were mostly within the range of measurement error (n = 35). FCM-AOT variables, in particular %Abnormal alpha(t), displayed significant correlations with motility (r = -0.557), vitality (r = -0.469) and morphology (r = -0.464, n = 201), which are similar in magnitude to those existing between the standard semen variables. Surprisingly, no correlation was found between %Abnormal alpha(t) and the microscopic acridine orange test (M-AOT) (n = 185), suggesting the FCM results are sensitive to a different aspect of sperm quality. In summary, this study confirms that although not totally independent of standard semen analysis or the M-AOT, it is found to be a robust, sensitive and reproducible measure of semen quality, representative of the individual.
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http://dx.doi.org/10.1016/s1472-6483(10)60923-6DOI Listing
April 2004

Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line.

Asian J Androl 2004 Mar;6(1):3-13

Department of Obstetrics and Gynaecology, University of Melbourne, 320 Victoria Parade, East Melbourne 3002 VIC, Australia.

Aim: To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests.

Methods: The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3 proteins individually and together by co-expression. Presence of these proteins in the culture medium and cell lysate was assessed by Western blotting analysis. The effect of the recombinant proteins on the human AR was assessed.

Results: RhZP2 and rhZP3 were secreted into the culture medium, whereas rhZP1 was found only in the cell lysate. Interestingly, when all zona pellucida proteins were co-expressed in the same cells, rhZP1 was also secreted into the culture medium. However, despite the presence of all three ZP proteins in sufficient concentration and evidence of heavy glycosylation on gel electrophoresis, biological activity to induce the AR was not observed.

Conclusion: RhZP1, rhZP2 and rhZP3 were successfully expressed in the human embryonic kidney cell line 293T. It appears that an interaction amongst these proteins may be required for release of rhZP1 from the cell. Although this approach is not satisfactory for producing active human ZP proteins, it makes a significant contribution to the understanding of the structural and functional characteristics of the ZP proteins.
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March 2004

High frequency of defective sperm-zona pellucida interaction in oligozoospermic infertile men.

Hum Reprod 2004 Feb;19(2):228-33

University of Melbourne Department of Obstetrics and Gynaecology, Reproductive Services, Royal Women's Hospital and Melbourne IVF, Victoria 3035, Australia.

Background: The ability of sperm to interact with the zona pellucida (ZP) plays a critical role during the process of human fertilization. The aim of this study is to determine frequency of defective sperm-ZP interaction in oligozoospermic infertile men.

Methods: Sperm-ZP binding assays and the ZP-induced acrosome reaction (AR) were performed in 72 infertile men with a sperm concentration <20 x 10(6)/ml. Oocytes that had previously failed to fertilize in a clinical IVF programme were used for the tests. Motile sperm (2 x 10(6)/ml) selected by swim-up from each semen sample were incubated with four oocytes for 2 h. The number of sperm bound per ZP and the ZP-induced AR were assessed. Under these conditions, an average of < or =40 sperm bound/ZP was defined as low sperm-ZP binding and a ZP-induced AR < or =16% was defined as low ZP-induced AR.

Results: In the 72 oligozoospermic men, 28% (20/72) had low sperm-ZP binding. Of those with normal sperm-ZP binding, 69% (36/52) had low ZP-induced AR. Overall, 78% (56/72) had either low ZP-binding or normal ZP binding but low ZP-induced AR. This means that only 22% (16/72) had both normal sperm-ZP binding and normal ZP-induced AR.

Conclusion: Oligozoospermic men have a very high frequency of defective sperm-ZP interaction, consistent with their low natural fertility or low fertilization rate in conventional IVF. Infertile couples with oligozoospermic semen should be treated by ICSI rather than by conventional IVF.
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http://dx.doi.org/10.1093/humrep/deh067DOI Listing
February 2004

Normal range and variation of the zona pellucida-induced acrosome reaction in fertile men.

Fertil Steril 2003 Aug;80(2):384-9

Department of Obstetrics and Gynaecology, University of Melbourne, Carlton, Victoria.

Objective: To investigate the normal range of the zona pellucida (ZP)-induced acrosome reaction (AR) in fertile men.

Design: A study of normative data.

Setting: Academic research and teaching tertiary hospital.None.Fertile men (n = 111) who had children or whose female partner was currently pregnant. MAIN OUTCOME MEASURE (S): Semen analysis, sperm-ZP binding, and the ZP-induced AR.

Result(s): There was wide range for the ZP-induced AR: 20%-98% with a mean of 48%. The ZP-induced AR was significantly correlated with sperm concentration and normal morphology. The ZP-induced AR was significantly lower in men with sperm concentration 60 x 10(6)/mL. Similarly, men with normal sperm morphology >or=15% had significantly higher ZP-induced AR than men with normal morphology <15%.

Conclusion(s): There is a wide range of the ZP-induced AR in fertile men that is correlated with sperm concentration and normal morphology.
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http://dx.doi.org/10.1016/s0015-0282(03)00603-4DOI Listing
August 2003

Frequency of defective sperm-zona pellucida interaction in severely teratozoospermic infertile men.

Hum Reprod 2003 Apr;18(4):802-7

University of Melbourne Department of Obstetrics and Gynaecology, Reproductive Services, Royal Women's Hospital and Melbourne IVF, Victoria 3035, Australia.

Background: The frequency of defective sperm-zona pellucida (ZP) interaction in teratozoospermic infertile men was investigated.

Methods: Sperm-ZP binding and the ZP-induced acrosome reaction (ZPIAR) were performed in 125 infertile men with <5% of their sperm with normal morphology (strict criteria), but with a sperm count > or =20x10(6)/ml and total motility >30% in semen. Oocytes that failed to fertilize in clinical IVF were used for the tests. Four oocytes were incubated for 2 h with 2x10(6)/ml motile sperm selected by swim-up. The number of sperm bound per ZP and ZPIAR were assessed. Under these conditions, an average < or sperm bound per ZP was defined as poor sperm-ZP binding, and a ZPIAR < or = was defined as low ZPIAR.

Results: Among 125 teratozoospermic men, 31% (39/125) had poor sperm-ZP binding. Of those without poor ZP binding, 48% (41/86) had low ZPIAR. Some 64% (28/44) with sperm counts between 20 and 60x10(6)/ml had low ZPIAR. Only 36% (45/125) had normal sperm-ZP binding and ZPIAR.

Conclusions: Defective sperm-ZP interaction was present in 64% of teratozoospermic infertile men: 31% had defective sperm-ZP binding, and 33% low ZPIAR. The frequency of low ZPIAR was higher in men with sperm counts between 20-60x10(6)/ml.
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http://dx.doi.org/10.1093/humrep/deg164DOI Listing
April 2003

Disordered zona pellucida-induced acrosome reaction and failure of in vitro fertilization in patients with unexplained infertility.

Fertil Steril 2003 Jan;79(1):74-80

University of Melbourne Department of Obstetrics and Gynecology, Reproductive Services, Royal Women's Hospital and Melbourne IVF, Melbourne, Victoria, Australia.

Objective: To determine the relationship between the zona pellucida (ZP)-induced acrosome reaction (AR) and fertilization rate and pregnancy rate in standard IVF and the frequency of disordered ZP-induced AR (DZPIAR) in patients with unexplained infertility.

Design: Prospective study.

Setting: Academic research and teaching tertiary hospital.

Patients: Patients with unexplained infertility with normal semen analysis.

Intervention: None.

Main Outcome Measure: Semen analysis, the ZP-induced AR, and measurements of fertilization rate and pregnancy rate with in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI).

Result(s): A statistically significant correlation was found for the ZP-induced AR and fertilization rate with standard IVF (n = 65). Patients with DZPIAR (ZP-induced AR of 16%, who had an average fertilization rate of 61%. The sensitivity and specificity of DZPIAR for prediction of IVF rates <30% and >or=30% were 80% and 86%, respectively. Of 260 patients screened, the frequency of DZPIAR was 29%. Ten patients with DZPIAR had an average fertilization rate of 15% and no pregnancy with initial IVF cycles, and a fertilization rate of 61% (with three live-birth pregnancies) with subsequent ICSI cycles. Another 33 patients with DZPIAR were treated with ICSI alone, with an average fertilization rate of 71% and a live-birth pregnancy rate of 17% per embryo transfer. Sixteen of the patients had live-birth pregnancies (including one set of twins) after undergoing an average of 3.2 embryo transfers.

Conclusion(s): Patients with DZPIAR have a low or zero fertilization rate with standard IVF but high fertilization and pregnancy rates with ICSI. Up to 29% of patients with unexplained infertility with normal semen analysis may have this condition, which should be diagnosed and treated with ICSI rather then standard IVF.
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http://dx.doi.org/10.1016/s0015-0282(02)04555-7DOI Listing
January 2003

Phorbol myristate acetate induces ruffling of the acrosome of human sperm.

Fertil Steril 2002 Jul;78(1):128-36

Department of Obstetrics and Gynaecology, University of Melbourne, Melbourne, Royal Women's Hospital, Victoria, Australia.

Objective: To determine the effect of phorbol myristate acetate (PMA) on human acrosome morphology and the acrosome reaction.

Design: Controlled experiments on sperm and unfertilized oocytes from volunteers.

Setting: Academic research and teaching tertiary hospital.

Patient(s): Sperm samples were from normospermic men and unfertilized oocytes from IVF patients.

Main Outcome Measure(s): Acrosome morphology was assessed by using transmission and scanning electron microscopy. The acrosome reaction was assessed by using fluorescein-labeled Pisum sativum agglutinin.

Result(s): PMA induced acrosome ruffling, indicated by a marked wavy appearance. A significant correlation was found between PMA-induced ruffling and PMA enhancement of the zona pellucida-induced acrosome reaction. Protein kinase C inhibitors bisindolylmalemide I and sangivamycin had no effect on PMA-induced acrosomal ruffling, but actin polymerization inhibitors cytochalasin B and cytochalasin D significantly decreased PMA-induced acrosomal ruffling. In contrast, bisindolylmalemide I, sangivamycin, cytochalasin B, and cytochalasin D significantly decreased both the zona pellucida-induced acrosome reaction and the PMA enhancement of the zona pellucida-induced acrosome reaction.

Conclusion(s): PMA-induced acrosomal ruffling involves actin polymerization, possibly independent of conventional protein kinase C. Acrosomal ruffling is involved in the PMA augmentation of the zona pellucida-induced acrosome reaction.
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http://dx.doi.org/10.1016/s0015-0282(02)03166-7DOI Listing
July 2002