An evaluation of embryo characteristic score to select the most favorable embryo for transfer

The need for establishing a greater knowledge of embryo quality variables is needed. Thus constructing reliable scoring systems is becoming increasingly more evident. This may result in selection of the appropriate embryo and reducing the number of embryos transferred.This paper was initially delivered at the Annual Congress of the Biological Sciences Division of the South African Academy for Science and Art, ARC-Plant Protection Research Institute, Roodeplaat, Pretoria, South Africa on 01 October 2010.http://www.satnt.ac.zaam2014ay201

Assisted reproduction laboratory cost-drivers in South Africa : value, virtue and validity

An overview is given on selected cost-drivers within an assisted reproduction technology (ART) laboratory, such as procedural costs; sperm preparations; laboratory supplies including embryo culture media and cryopreservation. Depending on the nature of an ART unit, i.e. private vs. public/tertiary, the structure of the unit will differ with regards to costs, services offered, and general patient population. ART laboratory equipment, culture media and disposables are imported from various parts of the world to South Africa. Costs will be influenced by the choice of ART diagnostic and therapeutic techniques, disposables and devices, whereby laboratory costs can escalate to near 50% of ART fees payable in the private sector. The ultimate goal of an ART treatment should be to achieve a healthy singleton as cost effectively as possible, especially in a developing country.This manuscript stemmed from an invited lecture presented at 35th National Congress of the South African Society of Obstetricians & Gynaecologists South African Society of Obstetricians and Gynaecologists (SASOG) in May 2012.http://www.ogf.co.z

Prevention of infections in an assisted reproduction laboratory in a developing country

The aim of the current paper is to discuss prevention of infections in a public sector laboratory orientated assisted reproduction treatment setting. Alternative viewpoints on resource management, semen decontamination for the human immunodeficiency virus type-1 (HIV-1) seropositive male and the subsequent choice of ART procedures, are addressed.This paper was initially delivered at the Annual Congress of the Biological Sciences Division of the South African Academy for Science and Art, ARC-Plant Protection Research Institute, Roodeplaat, Pretoria, South Africa on 01 October 2010.http://www.satnt.ac.zaam2014ay201

A patient journey of understanding – navigating the avenues of assisted reproductive technologies : three case studies

A study designed to determine what type of patients may attend a government-subsidized infertility clinic, the patients’ understanding of medical terms, jargon, sexually transmitted diseases and the level of stressors patients experience during an infertility workup.This paper was initially delivered at the Annual Congress of the Biological Sciences Division of the South African Academy for Science and Art, ARC-Plant Protection Research Institute, Roodeplaat, Pretoria, South Africa on 01 October 2010.http://www.satnt.ac.zaam201

Mitochondrial transfer : implications for assisted reproductive technologies

The use of mitochondrial transfer as a clinic procedure is drawing closer to reality. Here we provide a detailed overview of mitochondrial transfer techniques – both established and recent – including pronuclear, spindle, ooplasmic and blastomere transfer. Reasons as to why some techniques are more suitable for the prevention of mitochondrial DNA disease than others, as well as the advantages and disadvantages of each methodology, are discussed. The possible clinical introduction of these techniques has raised concerns about the adverse effects they may have on resultant embryos and offspring. Success rates of each technique, embryo viability and developmental consequences post mitochondrial transfer are addressed through analysis of evidence obtained from both animal and human studies. Counterarguments against potential mitochondrial-nuclear genome incompatibility are also provided. Additional clinical applications ofmitochondrial transfer techniques are discussed. These include the rescue or enhancement of fertility in women of advanced maternal age or those suffering from diabetes. An alternative to using mitochondrialDNA transfer for germline therapies is the therapeutic use of somatic cell nuclear transfer for the generation of personalised stem cells. Although ethically challenging, this method could offer patients already suffering from mitochondrial DNA diseases a novel treatment option.This work was supported by grants from the South African Medical Research Council Flagship Awards Project SAMRC-RFA-UFSP-01-2013/ STEM CELLS, the SAMRC Extramural Unit for Stem Cell Research and Therapy and the Institute for Cellular and Molecular Medicine of the University of Pretoria.http://www.elsevier.com/locate/atgam2017ImmunologyObstetrics and Gynaecolog

Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel tube insert

The occurrence of bacteria in sperm samples intended for in vitro fertilization, can compromise the outcome of assisted reproductive techniques. Effective semen processing procedures should therefore be implemented to remove bacteria from semen. Unfortunately, technique failure does occur whereby bacteria can be found in processed sperm preparations. To improve the effectiveness of semen processing, a novel centrifuge tube insert was developed to facilitate the layering of density gradients and semen, and to prohibit the reinfection of purified sperm pellets. The purpose of this study was to: 1) determine the prevalence and type of bacteria present in semen of patients participating in the Unit’s assisted reproduction program, and 2) evaluate the effectiveness of density gradient centrifugation with the novel tube insert, for the elimination of bacteria and yeast from spiked human semen samples. A survey in 2007-2010 indicated that 50% of semen samples were found to have positive bacterial cultures. Semen processing by means of density gradient centrifugation with the novel tube insert, eliminated significantly more in vitro derived (spiked) bacteria and yeast from semen compared to processing without the insert (P<0.004). Therefore, it is highly recommended that the centrifuge tube insert, ProInsertTM, be incorporated into assisted reproductive programs.Medical Research Council (MRC)http://www.blackwell-synergy.com/loi/an

Large volume cryoprotectant-free vitrification : an alternative to conventional cryopreservation for human spermatozoa

Vitrification is a simple and cost-effective method for the storage of human spermatozoa without the use of conventional cryoprotectants, by plunging the sperm suspension directly into liquid nitrogen. As a result, solidification of living cells without the formation of ice crystals is achieved during cooling. This study aimed to compare cryoprotectant-free vitrification to conventional cryopreservation protocols. Semen samples (n = 35) were collected from patients seeking diagnostic assistance at the Reproductive and Endocrine Unit at Steve Biko Academic Hospital. Samples were processed using a discontinu-ous density-gradient centrifugation method. Washed samples were split into two aliquots and cryopreserved either by means of cryoprotectant-free vitrifica-tion (sucrose + 1% albumin) or conventional slow freezing (TEST-yolk buf-fer). Post-thawing, the sperm motion parameters, mitochondrial membrane potential (Dwm) and DNA fragmentation were compared between the two groups. No significant differences were observed in the sperm motility parame-ters (P > 0.05). Significantly higher percentages of Dwm (11.99% 4.326%versus 6.58% 1.026%; P < 0.001) and lower percentages of DNA fragmenta-tion (2.79% 1.017% versus 3.86% 1.38%; P < 0.01) were observed when comparing cryoprotectant-free vitrification to conventional cryopreservation. Cryoprotectant-free vitrification is a rapid and promising alternative to conventional methods resulting in good-quality spermatozoa post-thaw.Research Committee of the Faculty of Health Sciences (RESCOM), University of Pretoria, Merck Serono (Pty) Ltd and the National Research Foundation.http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1439-02722016-06-30hb201

Treatment of human sperm with serine protease during density gradient centrifugation

PURPOSE Seminal pathogens can bind specifically or nonspecifically to spermatozoa, rendering semen decontamination procedures ineffective, whereby vertical or horizontal transmission of the infection could occur. Serine proteases have been demonstrated to effectively inactivate viruses and to break pathogen-sperm bonds. However, the addition of a protease to density gradient layers during semen processing could negatively impact on sperm parameters. This study investigated the effect of the addition of a recombinant, human-sequence protease (rhProtease) on sperm parameters during density gradient centrifugation. METHODS (i) Pooled semen samples (n09) were split and processed by density gradient centrifugation, with the top density layers supplemented, or non-supplemented with rhProtease at three different concentrations (diluted 2, 10 and 20 times). Sperm parameters were then analysed by flow cytometry and computer-assisted semen analyses. (ii) Semen samples (n05) were split and similarly processed using PureSperm® Pro, with rhProtease in the 40 % density gradient layer, or standard PureSperm® not supplemented with rhProtease (Nidacon, International) respectively. The Hemizona assay was then utilized to compare sperm-zona binding post processing. RESULTS Evaluation of sperm parameters indicated that rhProtease did not, at any of the tested concentrations, have an impact on (i) mitochondrial membrane potential, vitality, motility, or (ii) zona binding potential. CONCLUSION We report that the addition of rhProtease to density gradients is a non-detrimental approach that could improve the effectiveness of semen processing for the elimination of seminal pathogens, and benefit assisted reproduction outcome.The MRChttp://www.springerlink.com/content/104689

Semen decontamination for the elimination of seminal HIV-1

The risk of human immunodeficiency virus (HIV) transmission to the female partner, or potential offspring of an HIV-1 infected man can be reduced using semen decontamination procedures before assisted reproductive treatment (ART). The objective of this study was to determine the efficiency of decontaminating semen samples (n = 186) from 95 HIV-1 sero-positive patients. Aliquots of neat semen were submitted for viral validation by qualitative and quantitative polymerase chain reaction. Semen samples were processed by density gradient centrifugation in combination with a ProInsert™ tube after which aliquots of the processed sperm samples were analysed for the presence of HIV-1. Fifty-four percent of all tested neat semen samples tested positive for HIV-1 DNA, RNA or both (13.4%, 11.3% and 29.0%, respectively). From a total of 103 processed sperm samples that were submitted for viral validation, two samples tested positive for HIV-1 DNA and none for RNA. In conclusion, semen processing with the ProInsert™ followed by viral validation of processed sperm samples should be carried out when providing ART to couples where the male partner is HIV-1 sero-positive.Medical Research Council and National Research Foundation (85821/N00414).http://www.rbmonline.com/hb201

Infertility in Gabon : a survey to determine diagnostic and medical support to patients

A survey among gynaecologists practising in Libreville was conducted to ascertain the level of infertility assistance available, and the feasibility of establishing an intrauterine insemination programme in Gabon. This descriptive study invited gynaecologists (n=20) in both private and public hospitals, who are members at the Gabonese Society of Obstetricians Gynaecologists and Reproduction, to participate in the survey. Seventeen (85%) surveys were completed. The information obtained indicated that each gynaecologist consulted with more than fifty patients monthly, and nearly half (45%) of these consultations were infertility related. Male patients were referred to four different pathology laboratories in Libreville for basic semen analyses (without microbiology testing). Nearly 65% of the respondents referred female patients for further infertility treatments elsewhere. Approximately one-third of all couples were unable to access additional medical assistance. This survey can be viewed as motivation for health policymakers to initiate discussions to improve medical diagnostics and implement accessible fertility services in Gabon. Gynaecological expertise, together with a developed infrastructure in Libreville, could serve as an appropriate base for the advancement of reproductive treatment facilities.Une enquête auprès des gynécologues exerçant à Libreville a été menée pour déterminer le niveau d'assistance disponible en matière de la stérilité et la possibilité de mettre en place un programme d'insémination intra-utérine au Gabon. Cette étude descriptive a invité des gynécologues (n = 20) d‘hôpitaux privés et publics qui sont membres de la Société des obstétriciens, gynécologues et de la reproduction, à participer à l‘enquête. Dix-sept (85%) sondages ont été complétés. Les informations obtenues indiquent que chaque gynécologue a consulté plus de cinquante patients par mois et que près de la moitié (45%) de ces consultations étaient liées à l'infertilité. Les patients de sexe masculin ont été dirigés vers quatre laboratoires de pathologie différents à Libreville pour des analyses de base du sperme (sans tests microbiologiques). Près de 65% des interviewés ont envoyé des patientes ailleurs pour un traitement supplémentaire contre l'infertilité. Environ le tiers des couples n‘ont pas pu avoir accès à une assistance médicale supplémentaire. Cette enquête peut être considérée comme une motivation pour les décideurs en matière de santé d‘engager des discussions en vue d‘améliorer les diagnostics médicaux et de mettre en place des services de fertilité accessibles au Gabon. L'expertise gynécologique, associée à une infrastructure développée à Libreville, pourrait constituer une base appropriée pour le développement des installations de traitement de la reproduction.The Research Committee: School of Medicine, University of Pretoria (RESCOM) and the National Research Fund (NRF).https://journals.co.za/content/journal/ajrham2020Obstetrics and Gynaecolog