Aim： To investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3 （ART3） mRNA is expressed in human testes and, if so, whether the expression is cell type-specific. Methods： ART3 mRNA was deter...Aim： To investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3 （ART3） mRNA is expressed in human testes and, if so, whether the expression is cell type-specific. Methods： ART3 mRNA was determined in human testes and sperm by reverse transcription-polymerase chain reaction （RT-PCR）. The glycosylphosphatidylinositol linkage of ART3 was shown by treating ART3-transfected HEK-293-T cells with phospholipase C. Fluorescent activated cell sorter （FACS）-analyses were used to detect ART3 on mature spermatozoa and immunohistological studies to detect the protein in testes. Results： ART3 protein was shown to be present in testes. It was found on spermatocytes only. It was absent from spermatogonia, spermatids and spermatozoa. The absence of ART3 from spermatozoa was confirmed by FACS-analysis. ART3 protein was detected neither within a seminoma nor on Leydig cells. Conclusion： Here we show for the first time that ART3 protein is expressed in testes in particular on spermatocytes, indicating that ART3 exerts a specific function only required at a particular stage of spermatogenesis.展开更多
Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the eff...Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.展开更多
The aim of the study was to determine the degree of compliance of Polish laboratories with World Health Organization （WHO） recommendations, with regard to semen analysis methodology. A survey requesting information ...The aim of the study was to determine the degree of compliance of Polish laboratories with World Health Organization （WHO） recommendations, with regard to semen analysis methodology. A survey requesting information about methods of semen analysis was distributed to employees of 55 laboratories. Respondents who had participated in external seminological workshops （31%） were termed certified respondents （CR）, the remaining （69%）--non-certified respondents （NCR）. Only one laboratory （6%） in the CR group and none in the NCR were compliant with WHO guidelines for methods and equipment used to evaluate seminal volume, sperm motility, concentration, vitality and morphology. Most problems were of volume measurement （weighing method was reported by 17% of CR and 10% of NCR） and staining method for sperm morphology （Papanicolau or Diff-Quik were found in 33% of CR and 23% of NCR）. A three- or four-point grading of sperm motility was used by the majority of respondents; however, 17% of CR and 37% of NCR did not use a laboratory counter to tally spermatozoa. Although a haemocytometer method was used by 80% of laboratories in each group, the improved Neubauer chamber was used only by 42% of CR and 19% of NCR. In each group, 24% of laboratories did not perform a vitality test. Procedural errors and the interchangeable utilization of two or even three methods to analyse a given parameter was observed in both groups. The results indicate a need for standardisation of the methods and continuous, unified training in semen analysis in Polish laboratories.展开更多
It is estimated that in 2010, 1 in every 250 adults will be a childhood cancer survivor. Today, oncological surgery, radiotherapy and chemotherapy achieve relatively high rates of remission and long-term survival, yet...It is estimated that in 2010, 1 in every 250 adults will be a childhood cancer survivor. Today, oncological surgery, radiotherapy and chemotherapy achieve relatively high rates of remission and long-term survival, yet are often detrimental to fertility. Quality of life is increasingly important to long-term survivors of cancer, and one of the major quality-of-life issues is the ability to produce and raise normal children. Developments in the near future in the emerging field of fertility preservation in cancer survivors promise to be very exciting. This article reviews the published literature, discusses the effects of cancer treatment on fertility and presents the options available today thanks to advances in assisted-reproduction technology for maintaining fertility in male and female patients undergoing this type of treatment. The various diagnostic methods of assessing the fertility potential and the efficacy of in vitro fertilization （IVF） after cancer treatment are also presented.展开更多
The introduction of designer oestrogens as a treatment modality in hormone replacement in women has invited toconsider the concept of compounds with selective androgenic effects for male hormone replacement therapy. T...The introduction of designer oestrogens as a treatment modality in hormone replacement in women has invited toconsider the concept of compounds with selective androgenic effects for male hormone replacement therapy. The fullspectrum of the actions of testosterone may not be necessary of even undesired for certain indications for testosteronetreatment. To define for what indications certain androgenic properties are desired and undesired more insight in basicandrogen (patho)physiology is required. There is convincing evidence that aromatization of androgenic compounds tooestrogens might be an advantage for maintenance of bone mass and it might also mitigate negative effects of androgenson biochemical parameters of cardiovascular risks; the potentially negative effects of oestrogens on prostate pathology inageing men needs further elucidation. While the role of dihydro-testosterone (DHT) for the male sexual differentiationand for pubertal sexual maturation is evident, its role in mature and ageing males seems less significant or may even beharmful. It is, however, of note that a negative effect of DHT on prostate pathophysiology is certainly not proven.For male contraception a progestational agent with strong androgenic properties might be an asset. For most of theandrogenic actions the critical levels of androgens are not well established. The latter is relevant since the large amountof androgen molecules required for its biological actions (as compared to oestrogens)is an impediment in androgenreplacement modalities. There may be room for more biopotent androgens since delivery of large amounts of androgenmolecules to the circulation poses problems for treatment modalities.展开更多
Infertility is a common condition experienced by many men and women, and treatments are expensive. The World Health Organization and American Society of Reproductive Medicine define infertility as a disease, yet priva...Infertility is a common condition experienced by many men and women, and treatments are expensive. The World Health Organization and American Society of Reproductive Medicine define infertility as a disease, yet private companies infrequently offer insurance coverage for infertility treatments. This is despite the clear role that healthcare insurance plays in ensuring access to care and minimizing the financial burden of expensive services. In this review, we assess the current knowledge of how male infertility care is covered by insurance in the United States. We begin with an appraisal of the costs of male infertility care, then examine the state insurance laws relevant to male infertility, and close with a discussion of why insurance coverage for male infertility is important to both men and women. Importantly, we found that despite infertility being classified as a disease and males contributing to almost half of all infertility cases, coverage for male infertility is often excluded from health insurance laws. Excluding coverage for male infertility places an undue burden on their female partners. In addition, excluding care for male infertility risks missing opportunities to diagnose important health conditions and identify reversible or irreversible causes of male infertility. Policymakers should consider providing equal coverage for male and female infertility care in future health insurance laws.展开更多
文摘Aim： To investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3 （ART3） mRNA is expressed in human testes and, if so, whether the expression is cell type-specific. Methods： ART3 mRNA was determined in human testes and sperm by reverse transcription-polymerase chain reaction （RT-PCR）. The glycosylphosphatidylinositol linkage of ART3 was shown by treating ART3-transfected HEK-293-T cells with phospholipase C. Fluorescent activated cell sorter （FACS）-analyses were used to detect ART3 on mature spermatozoa and immunohistological studies to detect the protein in testes. Results： ART3 protein was shown to be present in testes. It was found on spermatocytes only. It was absent from spermatogonia, spermatids and spermatozoa. The absence of ART3 from spermatozoa was confirmed by FACS-analysis. ART3 protein was detected neither within a seminoma nor on Leydig cells. Conclusion： Here we show for the first time that ART3 protein is expressed in testes in particular on spermatocytes, indicating that ART3 exerts a specific function only required at a particular stage of spermatogenesis.
文摘Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.
文摘The aim of the study was to determine the degree of compliance of Polish laboratories with World Health Organization （WHO） recommendations, with regard to semen analysis methodology. A survey requesting information about methods of semen analysis was distributed to employees of 55 laboratories. Respondents who had participated in external seminological workshops （31%） were termed certified respondents （CR）, the remaining （69%）--non-certified respondents （NCR）. Only one laboratory （6%） in the CR group and none in the NCR were compliant with WHO guidelines for methods and equipment used to evaluate seminal volume, sperm motility, concentration, vitality and morphology. Most problems were of volume measurement （weighing method was reported by 17% of CR and 10% of NCR） and staining method for sperm morphology （Papanicolau or Diff-Quik were found in 33% of CR and 23% of NCR）. A three- or four-point grading of sperm motility was used by the majority of respondents; however, 17% of CR and 37% of NCR did not use a laboratory counter to tally spermatozoa. Although a haemocytometer method was used by 80% of laboratories in each group, the improved Neubauer chamber was used only by 42% of CR and 19% of NCR. In each group, 24% of laboratories did not perform a vitality test. Procedural errors and the interchangeable utilization of two or even three methods to analyse a given parameter was observed in both groups. The results indicate a need for standardisation of the methods and continuous, unified training in semen analysis in Polish laboratories.
文摘It is estimated that in 2010, 1 in every 250 adults will be a childhood cancer survivor. Today, oncological surgery, radiotherapy and chemotherapy achieve relatively high rates of remission and long-term survival, yet are often detrimental to fertility. Quality of life is increasingly important to long-term survivors of cancer, and one of the major quality-of-life issues is the ability to produce and raise normal children. Developments in the near future in the emerging field of fertility preservation in cancer survivors promise to be very exciting. This article reviews the published literature, discusses the effects of cancer treatment on fertility and presents the options available today thanks to advances in assisted-reproduction technology for maintaining fertility in male and female patients undergoing this type of treatment. The various diagnostic methods of assessing the fertility potential and the efficacy of in vitro fertilization （IVF） after cancer treatment are also presented.
文摘The introduction of designer oestrogens as a treatment modality in hormone replacement in women has invited toconsider the concept of compounds with selective androgenic effects for male hormone replacement therapy. The fullspectrum of the actions of testosterone may not be necessary of even undesired for certain indications for testosteronetreatment. To define for what indications certain androgenic properties are desired and undesired more insight in basicandrogen (patho)physiology is required. There is convincing evidence that aromatization of androgenic compounds tooestrogens might be an advantage for maintenance of bone mass and it might also mitigate negative effects of androgenson biochemical parameters of cardiovascular risks; the potentially negative effects of oestrogens on prostate pathology inageing men needs further elucidation. While the role of dihydro-testosterone (DHT) for the male sexual differentiationand for pubertal sexual maturation is evident, its role in mature and ageing males seems less significant or may even beharmful. It is, however, of note that a negative effect of DHT on prostate pathophysiology is certainly not proven.For male contraception a progestational agent with strong androgenic properties might be an asset. For most of theandrogenic actions the critical levels of androgens are not well established. The latter is relevant since the large amountof androgen molecules required for its biological actions (as compared to oestrogens)is an impediment in androgenreplacement modalities. There may be room for more biopotent androgens since delivery of large amounts of androgenmolecules to the circulation poses problems for treatment modalities.
文摘Infertility is a common condition experienced by many men and women, and treatments are expensive. The World Health Organization and American Society of Reproductive Medicine define infertility as a disease, yet private companies infrequently offer insurance coverage for infertility treatments. This is despite the clear role that healthcare insurance plays in ensuring access to care and minimizing the financial burden of expensive services. In this review, we assess the current knowledge of how male infertility care is covered by insurance in the United States. We begin with an appraisal of the costs of male infertility care, then examine the state insurance laws relevant to male infertility, and close with a discussion of why insurance coverage for male infertility is important to both men and women. Importantly, we found that despite infertility being classified as a disease and males contributing to almost half of all infertility cases, coverage for male infertility is often excluded from health insurance laws. Excluding coverage for male infertility places an undue burden on their female partners. In addition, excluding care for male infertility risks missing opportunities to diagnose important health conditions and identify reversible or irreversible causes of male infertility. Policymakers should consider providing equal coverage for male and female infertility care in future health insurance laws.
