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Featured researches published by I.M. Agbaje.


Systems Biology in Reproductive Medicine | 2008

Sperm DNA tests as useful adjuncts to semen analysis.

Sheena E.M. Lewis; I.M. Agbaje; Juan G. Alvarez

Male infertility has traditionally been diagnosed by microscopic assessment of concentration, motility and morphology of sperm in the ejaculate. Most laboratories use sperm isolated by various methods such as density gradient centrifugation to enrich for subpopulations of sperm believed to have greater fertilization potential. These tests are essential to provide the fundamental information on which clinicians base their initial diagnosis. However, in the clinical setting, tests with superior prognostic value are needed. Tests showing much promise are those determining sperm DNA integrity, particularly the Comet, TUNEL, and Sperm Chromatin Structure assays. Sperm nuclear DNA fragmentation has been positively correlated with lower fertilization rates in IVF, impaired implantation rates, an increased incidence of abortion and disease in offspring, including childhood cancer. The mitochondrial genome of sperm has also been shown to be a sensitive marker of sperm health. Although the usefulness of these tests is recognized, insufficient resources have been available to develop standardized tests and protocols that could lead to universally accepted clinical thresholds. Associated with the lack of useful prognostic tests is the lack of improvement in assisted conception success rates despite thirty years of worldwide use. International collaborations should be initiated to develop agreed protocols and establish clinical thresholds.


Mutagenesis | 2008

Using the alkaline comet assay in prognostic tests for male infertility and assisted reproductive technology outcomes

Sheena E.M. Lewis; I.M. Agbaje

Infertility affects one in six couples in Europe during their reproductive years with dysfunctional sperm being one of the most common causes. Conventional semen analysis has proven variable and lacking in prognostic value so, over the past decade, more useful molecular fertility biomarkers have been explored. Among the tests showing most promise are those measuring sperm DNA quality. Sperm DNA damage has been closely associated with numerous indicators of reproductive health, including, fertilization, embryo quality, implantation, spontaneous abortion and childhood diseases. It therefore has great potential as a prognostic test for assisted reproductive treatment (ART), when couples are presenting with male infertility. Unlike somatic cells, sperm have a unique tightly compacted chromatin structure. Our group has modified the alkaline comet assay for use with sperm. Sperm DNA also differs from somatic cells in its high susceptibility to oxidative damage; this is largely due to the presence of abundant polyunsaturated fatty acids acting as substrates for reactive oxygen species (ROS) and its lack of repair mechanisms. Consequently, the effects of ROS and antioxidant protection on sperm DNA fragmentation have been widely investigated. In this review, the relationship between actual sperm DNA damage as determined by the alkaline comet assay and potential DNA damage as measured by DNA adduct testing will also be examined and the potential of routine clinical practices such as cryopreservation and prolonged incubation to induce further DNA damage was investigated. Finally, the usefulness of sperm DNA tests as prognostic markers and in particular, the opportunities and challenges provided by DNA testing in male fertility determination will be discussed.


Reproductive Biomedicine Online | 2008

Increased concentrations of the oxidative DNA adduct 7, 8-dihydro-8-oxo-2-deoxyguanosine in the germ-line of men with type 1 diabetes

I.M. Agbaje; Carmel McVicar; Bettina C. Schock; Neil McClure; A.B. Atkinson; Deirdre Rogers; Sem Lewise

The effects of diabetes mellitus on male reproductive health have not been clearly defined. A previous publication from this group reported significantly higher levels of nuclear DNA fragmentation and mitochondrial DNA deletions in spermatozoa from men with type 1 diabetes. This study compared semen profiles, sperm DNA fragmentation and levels of oxidative DNA modification in spermatozoa of diabetic and non-diabetic men. Semen samples from 12 non-diabetic, fertile men and 11 type 1 diabetics were obtained and subjected to conventional light microscopic semen analysis. Nuclear DNA fragmentation was assessed using an alkaline Comet assay and concentrations of 7,8-dihydro-8-oxo-2-deoxyguanosine (8-OHdG), an oxidative adduct of the purine guanosine, were assessed by high-performance liquid chromatography. Conventional semen profiles were similar in both groups, whilst spermatozoa from type 1 diabetics showed significantly higher levels of DNA fragmentation (44% versus 27%; P < 0.05) and concentrations of 8-OHdG (3.6 versus 2.0 molecules of 8-OHdG per 10(5) molecules of deoxyguanosine; P < 0.05). Furthermore, a positive correlation was observed between DNA fragmentation and concentrations of 8-OHdG per 10(5) molecules of deoxyguanosine (rs = 0.7, P < 0.05). The genomic damage evident in spermatozoa of type 1 diabetics may have important implications for their fertility and the outcome of pregnancies fathered by these individuals.


International Journal of Andrology | 2009

Advanced glycation end products accumulate in the reproductive tract of men with diabetes

Con Mallidis; I.M. Agbaje; Deirdre Rogers; Josephine V. Glenn; R. Pringle; A.B. Atkinson; Klaus Steger; Alan W. Stitt; Neil McClure

Light microscopic studies comparing sperm parameters show little association between diabetes and male fertility. However, with the introduction of new analytical techniques, evidence is now emerging of previously undetectable effects of diabetes on sperm function. Specifically, a recent study has found a significantly higher sperm nuclear DNA fragmentation in diabetic men. As advanced glycation end products (AGEs) are important instigators of oxidative stress and cell dysfunction in numerous diabetic complications, we hypothesized that these compounds could also be present in the male reproductive tract. The presence and localization of the most prominent AGE, carboxymethyl-lysine (CML), in the human testis, epididymis and sperm was determined by immunohistochemistry. Parallel ELISA and Western blot analyses were performed to ascertain the amount of CML in seminal plasma and sperm from 13 diabetic and nine non-diabetic subjects. CML immunoreactivity was found throughout the seminiferous epithelium, the nuclei of spermatogonia and spermatocytes, in the basal and principle cells cytoplasm and nuclei of the caput epididymis and on most sperm tails, mid pieces and all cytoplasmic droplets. The acrosomal cap, especially the equatorial band, was prominently stained in diabetic samples only. The amount of CML was significantly higher (p = 0.004) in sperm from non-diabetic men. Considering the known detrimental actions of AGEs in other organs, the presence, location and quantity of CML, particularly the increased expression found in diabetic men, suggest that these compounds may play a hitherto unrecognized role in male infertility.


Fertility and Sterility | 2009

The influence of type 1 diabetes mellitus on spermatogenic gene expression

Con Mallidis; I.M. Agbaje; Jason O'Neill; Neil McClure

Routine semen analysis found no differences in diabetic men; however, mRNA profiles showed changes in the expression of genes involved in oxidative stress.


International Journal of Andrology | 2010

Differences in mouse models of diabetes mellitus in studies of male reproduction

Jason O'Neill; Agnieszka Czerwiec; I.M. Agbaje; Josephine V. Glenn; Alan W. Stitt; Neil McClure; Con Mallidis

Diabetes Mellitus (DM) has been found to have subtle yet profound effects on the metabolic status of the testis, the expression of numerous spermatogenic genes and is associated with increased numbers of sperm with nuclear DNA damage. The precise mechanism causing these detrimental effects remains unknown. The presence of increased levels of the most prominent member (carboxymethyllysine - CML) of the advanced glycation end product adducts and their receptor (RAGE) in the reproductive tract of DM men has provided a new avenue for research. As there are suspicions that the antibiotic (streptozotocin - STZ) employed to induce DM is also capable of causing oxidative stress and DNA damage, we compared CML and RAGE levels in the reproductive tract and sperm nDNA status of STZ mice with the levels in the Ins(2Akita) mouse to determine which more closely mimics the situation described in the human diabetic. CML was observed in the testes, epididymes and sperm of all animals. Sperm from DM mice showed particularly strong CML immunolocalization in the acrosomal cap, the equatorial region and whenever present, cytoplasmic droplets. Although increased, the level of CML on the sperm of the STZ and Ins(2Akita) DM mice did not reach statistical significance. RAGE was present on the developing acrosome and epididymal sperm of all animals and in discrete regions of the epididymes of the DM models. Only the epididymal sperm of the Ins(2Akita) mice were found to have significantly increased (p < 0.0001) nDNA damage. The Ins(2Akita) mouse therefore appears to more accurately reflect the conditions found in the human and, as such, is a more representative model for the study of diabetes and glycations influence on male fertility.


Human Reproduction | 2007

Insulin dependant diabetes mellitus: implications for male reproductive function

I.M. Agbaje; Deirdre Rogers; Carmel McVicar; Neil McClure; A.B. Atkinson; Con Mallidis; Sheena E.M. Lewis


Human Reproduction | 2007

Distribution of the receptor for advanced glycation end products in the human male reproductive tract: prevalence in men with diabetes mellitus

Con Mallidis; I.M. Agbaje; Deirdre Rogers; Josephine V. Glenn; Stephen McCullough; A. Brew Atkinson; Klaus Steger; Alan W. Stitt; Neil McClure


Reproductive Biomedicine Online | 2011

Male diabetes mellitus and assisted reproduction treatment outcome

James Mulholland; Con Mallidis; I.M. Agbaje; Neil McClure


Reproduction | 2007

Advanced Glycation End Products (AGEs) accumulate in the Reproductive Tract of Men with Diabetes.

Con Mallidis; I.M. Agbaje; Deirdre Rogers; Josephine V. Glenn; R. Pringle; A.B. Atkinson; Klaus Steger; Alan W. Stitt; Neil McClure

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Neil McClure

Queen's University Belfast

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Deirdre Rogers

Queen's University Belfast

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Alan W. Stitt

Queen's University Belfast

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Carmel McVicar

Queen's University Belfast

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Jason O'Neill

Queen's University Belfast

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