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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

The effect of insulin, leptin and inflammatory cytokines on reproductive health and hypogonadism in males diagnosed with the metabolic syndrome

Leisegang, Kristian January 2013 (has links)
Philosophiae Doctor - PhD / The metabolic syndrome (MetS) is a collection of various metabolic, hormonal and immunological risk factors that cluster together, closely related to poorly understood phenomena such a hyperinsulinaemia (insulin resistance), hyperleptinaemia (leptin resistance), a low grade, systemic and chronic inflammation and, in males, hypogonadism. Infertility is increasing globally, and male factor infertility accounts for a large percentage of couples who are not able to conceive. The relationship between components of MetS and male reproductive health is not clear, and requires further investigation, as does the impact of MetS on male reproductive health in a case controlled study. The impact of hyperinsulinaemia, hyperleptinaemia and inflammatory cytokines on the male reproductive tract also requires investigation. Furthermore, it is hypothesised that these phenomena negatively impact steroidogenesis cascades. In order to investigate this, a case controlled study and TM3 Leydig cell culture experiments were designed.Participants were recruited from public advertisements, and screened for strict exclusion criteria, including acute or chronic inflammation, hormonal treatments, vasectomy and leukocytospermia (> 106/ml). Following clinical diagnostics, 78 males were either placed into a control group (CG) or the MetS group, with numerous parameters compared between them. Serum was assayed for routine risk markers including HDL cholesterol, triglycerides, glucose and C-reactive protein (CRP). Saliva was assayed for free testosterone and progesterone. Semen samples underwent semen analysis for ejaculation volume, sperm concentration and motility, vitality, morphology and leukocyte concentration, in addition to mitochondrial membrane potential (MMP) and DNA fragmentation (DF). Both serum and seminal fluid were further assayed for insulin, leptin, tumour necrosis factor-alpha (TNF ) and interleukins 1-beta (IL1 ), 6 (IL6) and 8 (IL8). Glucose was also assayed in seminal fluid. Separately, hCG stimulated TM3 Leydig cells were exposed to varying concentrations of insulin (0.01, 0.1, 1 & 10 pg/ml), TNF , IL1 , IL6 and IL8 (0.1, 1, 10 & 100 pg/ml) for 48 hours at optimal cell culture conditions. TM3 cell viability, protein concentration and testosterone and progesterone concentrations were assessed.XXII Results indicated that males in the MetS group (n=34) had significantly increased body mass index, waist circumference, blood pressure, triglycerides, glucose, and Creactive protein (CRP) with decreased HDL cholesterol, as compared to the CG. Furthermore, ejaculation volume, sperm concentration, total sperm count, progressive and total motility were significantly decreased in the MetS group, and sperm with abnormal MMP and DF were increased in this group. No difference was found for morphology. Serum and seminal insulin, leptin, TNF , IL1 , IL6 and IL8 were all significantly increased in the MetS group. Both testosterone and progesterone were also significantly decreased in the MetS group. Insulin increased testosterone and decreased progesterone in the TM3 cells. TNF , IL1 and IL6 all decreased testosterone and progesterone concentrations and TM3 cell viability. IL8 increased TM3 cell viability and decreased progesterone, will no effect on testosterone. These results suggest MetS is associated with decreased fertility potential in males. Furthermore, a significant increase in seminal insulin, leptin, TNF , IL1 , IL6 and IL8 suggests local reproductive tract inflammation in the absence of leukocytospermia. Strong correlations between serum and seminal insulin, leptin, TNF , IL1 , IL6 and IL8, as well as serum CRP, imply that these systemic phenomenons are related to the reproductive tract changes observed. Therefore, the underlying pathophysiology of MetS negatively affects male reproduction, in addition to general health and wellbeing. A decrease in progesterone and testosterone suggests a collapse in steroidogenesis cascades. Additionally, inflammation, increased leptin and insulin resistance likely contribute to this collapse in steroidogenesis based on TM3 cell culture experiments. These results provide novel avenues for further investigations.
2

Investigations on the in vitro effects of aqueous Eurycoma longifolia Jack extract on male reproductive functions

Erasmus, Nicolete January 2012 (has links)
<p>Eurycoma longifolia (Tongkat Ali / TA) is a Malaysian shrub used to treat various illnesses including male infertility. Considering that TA is also used to improve male fertility and no report&nbsp / regarding its safety has been published, this study investigated the effects of a patented, aqueous TA extract on various sperm and testicular functions. Materials and Methods This study&nbsp / encompasses two parts (part 1: on spermatozoa / part 2: on TM3-Leydig and TM4-Sertoli cells). Part 1: Semen samples of 27 patients and 13 fertile donors were divided into two groups,&nbsp / washed and swim-up prepared spermatozoa, and incubated with different concentrations of TA (1, 10, 20, 100, 2000 &mu / g/ml) for 1 hour at 37&deg / C. A sample without addition of TA served as control. After incubation with TA,&nbsp / the following parameters were evaluated: viability (Eosin-Nigrosin test), total and progressive motility (CASA), acrosome reaction (triple stain technique), sperm production of reactive oxygen&nbsp / species (ROS / dihydroethidium test / DHE), sperm DNA fragmentation (TUNEL assay) and mitochondrial membrane potential (&Delta / &psi / m) (Depsipher kit). Part 2: TM3-Leydig and TM4-Sertoli cells&nbsp / incubated with different concentrations of TA (0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50 &mu / g/ml) and control (without extract) for 48 and 96 hours. After incubation with TA, the following parameters were&nbsp / evaluated: viability (XTT), cell proliferation (protein assay), testosterone (testosterone ELISA test) and pyruvate (pyruvate assay). Results Part 1: For washed spermatozoa, significant&nbsp / dose-dependent trends were found&nbsp / for viability, total motility, acrosome reaction and sperm ROS production. However, these trends were only significant if the highest concentrations were included in the calculation. In the swim-up spermatozoa, ROS production of spermatozoa showed a biphasic relationship with its lowest percentage at 10 &mu / g/ml, yet, no significance could be&nbsp / observed (P=0.9505). No influence of TA could be observed for sperm DNA fragmentation nor &Delta / &psi / m.</p>
3

Investigations on the in vitro effects of aqueous Eurycoma longifolia Jack extract on male reproductive functions

Erasmus, Nicolete January 2012 (has links)
<p>Eurycoma longifolia (Tongkat Ali / TA) is a Malaysian shrub used to treat various illnesses including male infertility. Considering that TA is also used to improve male fertility and no report&nbsp / regarding its safety has been published, this study investigated the effects of a patented, aqueous TA extract on various sperm and testicular functions. Materials and Methods This study&nbsp / encompasses two parts (part 1: on spermatozoa / part 2: on TM3-Leydig and TM4-Sertoli cells). Part 1: Semen samples of 27 patients and 13 fertile donors were divided into two groups,&nbsp / washed and swim-up prepared spermatozoa, and incubated with different concentrations of TA (1, 10, 20, 100, 2000 &mu / g/ml) for 1 hour at 37&deg / C. A sample without addition of TA served as control. After incubation with TA,&nbsp / the following parameters were evaluated: viability (Eosin-Nigrosin test), total and progressive motility (CASA), acrosome reaction (triple stain technique), sperm production of reactive oxygen&nbsp / species (ROS / dihydroethidium test / DHE), sperm DNA fragmentation (TUNEL assay) and mitochondrial membrane potential (&Delta / &psi / m) (Depsipher kit). Part 2: TM3-Leydig and TM4-Sertoli cells&nbsp / incubated with different concentrations of TA (0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50 &mu / g/ml) and control (without extract) for 48 and 96 hours. After incubation with TA, the following parameters were&nbsp / evaluated: viability (XTT), cell proliferation (protein assay), testosterone (testosterone ELISA test) and pyruvate (pyruvate assay). Results Part 1: For washed spermatozoa, significant&nbsp / dose-dependent trends were found&nbsp / for viability, total motility, acrosome reaction and sperm ROS production. However, these trends were only significant if the highest concentrations were included in the calculation. In the swim-up spermatozoa, ROS production of spermatozoa showed a biphasic relationship with its lowest percentage at 10 &mu / g/ml, yet, no significance could be&nbsp / observed (P=0.9505). No influence of TA could be observed for sperm DNA fragmentation nor &Delta / &psi / m.</p>
4

Investigations on the in vitro effects of aqueous Eurycoma longifolia Jack extract on male reproductive functions

Erasmus, Nicolete January 2012 (has links)
Magister Scientiae (Medical Bioscience) - MSc(MBS) / Introduction: Eurycoma longifolia (Tongkat Ali; TA) is a Malaysian shrub used to treat various illnesses including male infertility. Considering that TA is also used to improve male fertility and no report regarding its safety has been published, this study investigated the effects of a patented, aqueous TA extract on various sperm and testicular functions. Materials and Methods: This study encompasses two parts (part 1: on spermatozoa; part 2: on TM3-Leydig and TM4-Sertoli cells). Part 1: Semen samples of 27 patients and 13 fertile donors were divided into two groups, washed and swim-up prepared spermatozoa, and incubated with different concentrations of TA (1, 10, 20, 100, 2000 μg/ml) for 1 hour at 37°C. A sample without addition of TA served as control. After incubation with TA, the following parameters were evaluated: viability (Eosin-Nigrosin test), total and progressive motility (CASA), acrosome reaction (triple stain technique), sperm production of reactive oxygen species (ROS; dihydroethidium test; DHE), sperm DNA fragmentation (TUNEL assay) and mitochondrial membrane potential (Δψm) (Depsipher kit). Part 2: TM3-Leydig and TM4-Sertoli cells incubated with different concentrations of TA (0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50 μg/ml) and control (without extract) for 48 and 96 hours. After incubation with TA, the following parameters were evaluated: viability (XTT), cell proliferation (protein assay), testosterone (testosterone ELISA test) and pyruvate (pyruvate assay). Results Part 1: For washed spermatozoa, significant dose-dependent trends were found for viability, total motility, acrosome reaction and sperm ROS production. However, these trends were only significant if the highest concentrations were included in the calculation. In the swim-up spermatozoa, ROS production of spermatozoa showed a biphasic relationship with its lowest percentage at 10 μg/ml, yet, no significance could be observed (P=0.9505). No influence of TA could be observed for sperm DNA fragmentation nor Δψm. Part 2: The viability rates and protein production of TM3-Leydig and TM4-Sertoli cells at 48-hour exposure to TA showed increases whereas at 96-hour incubation periods viability and protein production declined especially as from concentration 25 μg/ml TA. Similar results could be seen for TM4-Sertoli cells pyruvate production. The testosterone production at 48-hour exposure marginally increased (P=0.0580) at the highest (50 μg/ml) concentration of TA. However, at 96-hour exposure to TA the testosterone production significantly (P=0.0065) increased. It is also apparent that after 96 hours the concentration of testosterone has increased [12 x 10-4 ng/ml] when compared to 48-hour exposure [6 x 10-7ng/ml] of Tongkat Ali. Conclusion: Part 1: Results indicate that the Tongkat Ali extract has no deleterious effects on sperm functions at therapeutically used concentrations (<2.5 μg/ml). Part 2: The cytotoxic effect of TA are only presented at higher concentration from 25 μg/ml. TM3-Leydig cells appears to be more resilient than TM4-Sertoli cells in viability and protein production yet at prolonged periods of exposure it is detrimental. Testosterone production only increases after 96 hours exposure to TA.

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