Thesis (MMed)-- Stellenbosch University, 2013. / ENGLISH ABSTRACT: AIM: Review outcome of pregnancies following successful external cephalic version (ECV) for breech presentation at term, particularly the caesarian section (CS) rate. ECV is a safe procedure with a minimal cost implication that can reduce non-cephalic presentation at onset of labour at term. The outcome of pregnancies following successful ECV is certainly of interest. A meta analysis of studies done between 1997 and 2004, found that pregnancies after successful ECV at term were not the same as those with spontaneous cephalic presentations and was associated with a CS rate twice that in pregnancies with spontaneous cephalic presentations. The conclusion was that pregnancies after successful ECV should not be considered the same as normal pregnancies. In a matched retrospective analysis of CS risk after successful ECV, done in the USA, it was concluded that CS delivery and operative vaginal delivery rates following successful ECV, were not increased. To date there are no such studies in South Africa. METHODOLGY: A retrospective descriptive study was done to audit all successful ECV’s done at the Fetal Evaluation Clinic (FEC) of Tygerberg Academic Hospital. The electronic data from the FEC was searched for successful ECV patients. The facilities where these patients delivered were identified. The outcome of the pregnancies was determined from patient files and/or the labor registers. The relevant information of each patient was captured. All file reviews and data capturing was done by the principal investigator. RESULTS: A total of 78 patients were included in the study. The median age was 28.7 years with a range from 17 to 40 years, the median parity 1 and the range 0 to 6 and the median body mass index 27.2 and the range 18.2 to 45.0. The method of determining gestational age is known in 71 (91%) patients of whom 37 (52%) had an early ultrasound examination. The median gestational age at ECV was 37 weeks with the inter quartile range 36 to 38 weeks. The median ECV to delivery time was 2 weeks with the inter quartile range 1 to 4 weeks. Higher levels of care were required at time of delivery by 47 (60.3%) patients. Vaginal deliveries occurred in 49 patients and 29 (37.2%) had CS. The most common indications for CS were cephalo pelvic disproportion 8, fetal distress 6, reversion back to breech presentations 4 and other abnormal presentations 4 (2 face presentations and 2 transverse lies). The mean birth weight of the babies was 3360g and the range 2100 to 4655g. On comparing the groups that had vaginal deliveries and CS, only nulliparous patients had a significantly (p=0.02) higher risk for CS. CONCLUSIONS: Following successful ECV all patients need to be carefully followed up for possible reversion to breech presentation or transverse lie. Nulliparous and gravid 2 para 1 patients with a previous CS need to be delivered in hospitals with CS facilities. Further studies are required to assess whether successful ECV results in more face presentations. / AFRIKAANSE OPSOMMING: DOELWIT: Om die uitkoms van swangerskappe na suksesvolle eksterne kefaliese kerings (EKK) vir stuit presentasies op voltyd, spesifiek die keisersnit (KS) insidensie te bepaal. EKK is ‘n veilige prosedure wat teen minimale koste die nie-kefaliese presentasies op voltyd kan verminder. Die uitkoms van swangerskappe na suksesvolle EKK is van belang. ‘n Meta-analise van studies gedoen tussen 1997 en 2004 vind dat swangerskappe na suksesvolle EKK op voltyd nie dieselfde is vergeleke met spontane kefaliese presentasies nie en gepaard gaan met ‘n KS koers tweekeer hoër as dié met spontane kefaliese presentasies op voltyd. Die gevolgtrekking was dat swangerskappe na suksesvolle EKK nie as normale swangerskappe beskou moet word nie. In ‘n gepaarde retrospektiewe ontleding van die KS risiko wat in die VSA gedoen is, word gevind dat die KS en operatiewe vaginale verlossing koerse na suksesvolle EKK, nie verhoog is nie. Tot op hede is daar geen studies hieroor in Suid-Afrika gedoen nie. METODE: ’n Retrospektiewe beskrywende studie is gedoen om all suksesvolle EKK wat by die Fetale Evaluasie Kliniek (FEK) gedoen is te oudit. ‘n Elektroniese data soektog van suksesvolle EKK by die FEK is gedoen. Die instellings waar die pasiënte verlos is, is vasgestel. Die uitkoms van die swangerskappe is bepaal deur pasiënt lêers en/of die kraamregisters na te gaan. Die relevant inligting oor elke pasiënt is versamel. RESULTATE: ‘n Totaal van 78 pasiënte is by die studie ingesluit. Die mediane ouderdom was 28.7 jaar met ‘n reikwydte van 17 tot 40 jaar, die mediane pariteit was 1 met ‘n reikwydte van 0 tot 6 en die mediane liggaamsmassa indeks 27.2 met ‘n reikwydte van 18.2 tot 45.0. Die metode waavolgens swangerskapsduurte bepaal is, was bekend in 71 (91%) van pasiënte, waarvan 37 (52%) vroeë ultraklank ondersoeke gehad het. Die mediane swangerskapsduurte tydens die EKK was 37 weke met die interkwartiele interval 36 tot 38 weke. Die mediane EKK tot verlossing tydsverloop was 2 weke met die interkwartiele interval 1 tot 4 weke. Hoër vlakke van sorg was nodig ten tye van die verlossing by 47 (60.3%) van pasiënte. Van die pasiënte het 49 vaginale verlossings en 29 (37.2%) KS gehad. Die mees algemene indikasies vir KS was skedel-bekken disproporsie 8, fetale nood 6, terugkeer na stuitpresentasie 4 en abnormale presentasies 4 (2 aangesigsliggings en 2 transversliggings). Die gemiddelde geboorte gewig van die babas was 3360g en die reikwydte 2100 tot 4655g. Wanneer die groep wat vaginale verlossing en KS gehad het vergelyk word, het slegs nullipareuse pasiënte ‘n betekenisvolle (p=0.02) hoër risiko vir KS gehad. GEVOLTREKKING: Na suksesvolle EKK moet alle pasiënte noukeurig opgevolg word vir terugkeer na ‘n stuit presentasie of transversligging. Nullipareuse en gravida 2 para 1 pasiënte met ‘n vorige KS moet in hospitale met KS fasiliteite verlos word. Verdere studies is nodig om te bepaal of suksesvolle EKK meer aangesig presentasies tot gevolg het.

Thesis (MMed)-- Stellenbosch University, 2013. / ENGLISH ABSTRACT: Misoprostol is often used for the purpose of induction of labour. However, its effect on fetal heart rate has not been systematically studied. Objective To assess the effect of misoprostol on fetal heart rate parameters during induction of labour from 38 completed weeks in women with previous intrauterine death or postterm pregnancy. Study design A retrospective descriptive study of 127 women for a period of 18 months. Method Women who underwent induction of labour with misoprostol for either previous intrauterine death or postterm pregnancy at Tygerberg hospital were eligible. The selected process of induction of labour happened according to the departmental protocol. The primary outcomes were changes in fetal heart rate (variability, accelerations and decelerations) pre-and post-administration of misoprostol. Secondary outcomes were neonatal highcare or intensive care unit (ICU) admissions. Results There was no statistical difference in the mean fetal heart rate and baseline variability in relation to time recordings after administration of misoprostol. There were no statistically significant differences in the distribution of accelerations and decelerations in different time intervals before and after administration. There were more reactive patterns at all time intervals after the administration of misoprostol, but these differences did not quite reach statistical significance. In both study groups no neonatal complications or intensive care admissions were reported. Conclusion In the absence of contra indications, 50mcg of oral misoprostol can be given to mothers for induction of labour as no harmful fetal heart tracing abnormalities were found for 45 minutes; however large prospective randomized controlled trials are still needed to confirm effectiveness and evaluate further maternal and neonatal safety issues. Optimal dose and frequency also still need robust interrogation. Based on this thesis it does appear that misoprostol is probably not harmful to the fetus under these circumstances. / AFRIKAANSE OPSOMMING: Misoprostol word dikwels gebruik vir induksie van kraam. Die effek daarvan op fetale hartspoed is egter nie sistematies ondersoek nie. Doel Om die effek van misoprostol op fetale hartspoedparameters gedurende die induksie van kraam van 38 voltooide weke in vroue met vorige intra-uteriene dood or oortyd swangerskap te evalueer. Studei-ontwerp „n Retrospektiewe beskrywende studie van 127 vroue oor „n periode van 18 maande. Metode Vroue wat induksie van kraam met misoprostol ondergaan het vir of vorige intra-uteriene dood of oortyd swangerskap by Tygerberg Hospitaal is ingesluit. Die proses van induksie van kraam is volgens departementele protokol uitgevoer. Die primêre uitkomste was veranderinge in fetale hartspoed (variasie, versnellings en verstadigings) pre- en post-toediening van misoprostol. Neonatale hoësorg of intensiewe sorg toelatings was sekondêre uitkomste. Resultate Ons het geen statistiese verskille in gemiddelde fetale hartspoed en basislynvariasie in verhouding tot die tyd na toediening van misoprostol gevind nie. Daar was geen statisties betekenisvolle verskille in die verspreiding van versnellings en verstadigings in verskillende tydsintervalle nie. Daar was meer reaktiewe patrone gedurende alle tydsintervalle na die toediening van misoprostol, maar hierdie verskille was nie statisties betekenisvol nie. In beide studiegroepe was daar geen neonatale komplikasies of intensiewe sorg toelatings nie. Gevolgtrekking In die afwesigheid van kontra-indikasies kan 50 mcg misoprostol aan moeders toegedien word vir induksie van kraam aangesien geen skadelike fetale hartsped abnormaliteite gevind is nie. Groot prospektiewe gerandomiseerde gekontroleerde studies word steeds benodig om effektiwiteit te bevestig en om moederlike en fetale veiligheidskwessies verder te evalueer. Optimale dosis en frekwensie benodig ook robuuste ondersoek. Gebaseer op hierdie tesis kom dit voor of misoprostol waarskynlik nie skadelik vir die fetus onder hierdie omstandighede nie.

Thesis (MMed)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Background Rectovaginal endometriosis accounts for 5-10% of cases of endometriosis and constitutes one of the forms of deep infiltrating endometriosis. . Deep infiltrating endometriosis involving the bowel is most frequently encountered in the rectovaginal septum and is considered to be the most severe form of the disease and the most difficult to treat surgically owing to its invasive nature. There are currently no studies on this topic pertaining to a South African context. Study Objective To document the outcomes in 112 patients undergoing laparoscopic surgery for rectovaginal endometriosis. Methods A retrospective audit of 112 women undergoing laparoscopic surgery for rectovaginal endometriosis at Vincent Pallotti's Aevitas Fertility Clinic was undertaken. Eligibility was established by identifying women from a surgical database based on medical aid coding as well as a review of individual case notes. Patients were telephonically contacted to gather any missing information and to assess further outcomes. Design Classification Study number S11/11/036. This study was approved by the Health Research Ethics Committee at Stellenbosch University and was conducted according to ethical guidelines and principles of The International Declaration of Helsinki, South African Guidelines for Good Clinical Practice and the Medical Research Council (MRC) Ethical Guidelines for Research. Setting Vincent Pallotti’s Aevitas Reproductive Medicine Clinic Patients 112 consecutive patients suffering from rectovaginal endometriosis Interventions: Laparoscopic surgery for treatment of deep infiltrating, namely rectovaginal endometriosis Measurements and Main Results Primary outcome: Complications of laparoscopic surgery for rectovaginal endometriosis included one patient requiring a blood transfusion (0,9%), three cases of rectovaginal fistula (2,7%), two bowel injuries (1,8%)-detected and managed intra-operatively , one ureteric injury (0,9%), one pelvic abscess (0,9%) and the need for three urgent re-operations (2,68%). Secondary outcome: Of the 71 patients desiring fertility 39 (54,9%) fell pregnant of which 27 (69,2%) were spontaneous. Conclusion To our knowledge this is the first study assessing surgical outcomes in the management of deep infiltrating endometriosis from South Africa. These outcomes are in keeping with complication rates quoted in the international literature. Most of the surgery was performed using the shaving technique, in keeping with international trends, whilst fourteen cases required the performance of a segmental resection owing to extensive disease. In trained hands laparoscopic surgery is a valid management option in the management of rectovaginal endometriosis. / AFRIKAANSE OPSOMMING: Agtergrond Vyf tot tien persent van alle endometriose gevalle kan toegeskryf word aan rektovaginale endometriose. Dit word beskou as een van die vorme van diep infiltrerende endometriose. Diep infiltrerende endometriose van die derm kom meestal in die rektovaginale septum voor en word as die ernstigste vorm van die siekte beskou. Dit is die moeilikste om chirurgies te behandel weens sy indringende aard. Daar is tans geen studies beskikbaar oor hierdie onderwerp in die Suid-Afrikaanse konteks nie. Doel van die studie Om die uitkomste te dokumenteer van 112 pasiënte wat laparoskopiese chirurgie vir rektovaginale endometriose ondergaan het. Metodes 'n Retrospektiewe oudit is by Vincent Pallotti se Aevitas Fertiliteitskliniek gedoen van 112 vroue wat laparoskopiese chirurgie vir rektovaginale endometriose ondergaan het. Geskikte pasiënte is geïdentifiseer vanaf 'n chirurgiese databasis gebaseer op mediese kodering, sowel as vanaf 'n oorsig van pasiënt notas. Pasiënte is telefonies genader om ontbrekende inligting in te samel en verdere uitkomste te evalueer. Klassifikasie Ontwerp Studie nommer S11/11/036. Hierdie studie is deur die Gesondheids Navorsing Etiese Komitee van die Universiteit van Stellenbosch goedgekeur en uitgevoer volgens die etiese riglyne en beginsels van die Internasionale Verklaring van Helsinki, Suid-Afrikaanse Riglyne vir Goeie Kliniese Praktyk en die Mediese Navorsingsraad (MNR) se Etiese Riglyne vir Navorsing. Instelling Vincent Pallotti se Aevitas Reproduktiewe Medisyne Kliniek Pasiënte 112 agtereenvolgende pasiënte met rektovaginale endometriose. Ingrepe: Laparoskopiese chirurgie vir die behandeling van diep infiltrende, rektovaginale endometriose. Resultate Primêre uitkoms: Komplikasies van laparoskopiese chirurgie vir rektovaginale endometriose het ingesluit: een pasiënt wat 'n bloedoortapping benodig het (0,9%), drie gevalle van rektovaginale fistels (2,7%), twee dermbeserings (1,8%) - intraoperatief gediagnoseer en herstel, een ureter besering (0,9%), een bekkenabses (0,9%) en drie dringende herhaal operasies (2,68%). Sekondêre uitkoms: Van die 71 pasiënte wat fertiliteit verlang het: 39 (54,9%) het swanger geraak, waarvan 27 (69,2%) spontaan was. Gevolgtrekking Sover ons kennis strek, is dit die eerste Suid-Afrikaanse studie waar daar na die chirurgiese uitkomste in die behandeling van diep infiltrerende endometriose gekyk is. Hierdie uitkomste stem ooreen met internasionale literatuur in terme van komplikasie syfers. Die meeste van die operasies is uitgevoer met behulp van die skeer-tegniek, in ooreenstemming met internasionale tendense, terwyl veertien gevalle segmentele reseksies vereis het weens uitgebreide siekte. In goed opgeleide hande is die laparoskopiese behandeling van rektovaginale endometriose ‘n geldige behandelings opsie.

Thesis (MMed)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Introduction Pregnancies in women with a previous unexplained stillbirth may be jeopardized by increased antenatal surveillance and higher rates of induction of labour and caesarean delivery without clear evidence of benefit. Despite the fact that there have been no studies that adequately tested fetal benefit in routine induction of labour for a previous stillbirth, a policy of routine induction of labour at 38 weeks, with all the associated maternal, fetal and health-care associated costs, was in practice at Tygerberg Hospital for the past 30 years. This study aimed to investigate the safety of continuation of these pregnancies until term (≥39 weeks). Aims and Objectives To assess the clinical outcome and impact on the health service in a pregnancy with a previous unexplained intra-uterine demise (IUD) by routine induction of labour at term instead of at 38 weeks. Methodology This was a prospective observational study on the safety of a new hospital protocol which was introduced in 2012. The protocol extended the gestation for induction after a previous IUD from 38 weeks to term. The study population included all pregnant patients with a current singleton pregnancy, and a previous unexplained or unexplored (no data available) singleton fetal demise ≥24 weeks/500grams. All patients with a previous stillbirth in the metropolitan drainage area of Tygerberg Hospital are referred to Tygerberg for further care; and all referrals during 2012 were recruited for the study. Patients with known or recurrent risks for intra-uterine death were managed according to the relevant clinical condition and were excluded from the study. Results During the audit period, 306 patients with a previous intra-uterine fetal death were referred for further management. Of these, 161 had a clear indication for either earlier intervention or no intervention and were excluded from the protocol. Of the remaining 145 patients, 9 met exclusion criteria and there were 2 patients who defaulted. Forty-two of the study patients (with no known previous medical problems) developed complications during their antenatal course that necessitated a change in clinical management and earlier (<39 weeks) delivery. Of the remaining 92 patients in the audit, 47 (51%) went into spontaneous labour before their induction date. There were no intra-uterine deaths prior to delivery. Conclusions Careful follow up at a high risk clinic identifies new or concealed maternal or fetal complications in 29% of patients with a previous IUD and no obvious maternal or fetal disease in the index pregnancy. When all risks are excluded and the pregnancy allowed to progress to 39 weeks before an induction is offered, 51% will go into spontaneous labour. / AFRIKAANSE OPSOMMING: Inleiding Swangerskappe in vroue met vorige onverklaarbare stilgeboorte mag in gevaar gestel word deur meer intense voorgeboorte sorg en ‘n groter hoeveelheid induksies van kraam en keisersnitte sonder duidelike bewyse dat dit tot voordeel strek. Ten spyte van die feit dat daar geen studies is wat bewys het dat roetine induksie van kraam vir ‘n vorige stilgeboorte op 38 weke tot voordeel van die baba was nie, was ‘n beleid van roetine induksie van kraam op 38 weke, met al die geassosieërde moederlike en fetale risikos daaraan verbonde; asook die hoë gesondheidskostes, roetine praktyk in Tygerberg Hospitaal vir die afgelope 30 jaar. Hierdie studie het ten doel gehad om die veiligheid van voortsetting van hierdie swangerskappe tot voltyd (≥39 weke) te ondersoek. Doelwitte Om die kliniese uitkoms; asook die impak op gesondheidsdienste te evalueer in ‘n swanger vrou met n vorige onverklaarbare intra-uteriene sterfte; deur roetine induksie van kraam aan te bied op voltyd in plaas van 38 weke. Metodologie Hierdie was n prospektiewe kohort studie om die veiligheid van ‘n nuwe hospitaal protokol wat in 2012 geïmplimenteer is, te bepaal. Hierdie protokol het die gestasie tydperk van induksie van kraam van alle swanger pasiënte na ‘n vorige onverklaarbare stilgeboorte van 38 weke na voltyd verleng. Die studiepopulasie het alle swanger pasiënte met ‘n huidige enkelswangerskap en ‘n vorige onverklaarbare of onbekende (geen data beskikbaar) enkelvoudige fetale sterfte ≥24 weke/500gram, ingesluit. Alle pasiënte in die metropolitaanse dreineringsarea van Tygerberg Hospitaal met ‘n vorige stilgeboorte word na Tygerberg verwys vir verdere hantering, en alle verwysings gedurende 2012 was gewerf vir die studie. Pasiënte met bekende of herhalende risikofaktore vir ‘n intra-uteriene sterfte was hanteer volgens die relevante kliniese inligting en was uitgesluit by die studie. Resultate Drie-honderd-en-ses pasiënte met ‘n vorige intra-uteriene fetale sterfte was gedurende die oudit periode verwys vir verdere hantering. In 161 pasiënte was daar ‘n duidelike indikasie vir of vroeër intervensie of geen intervensie nie; en hulle was uitgesluit van die protokol. Van die oorblywende 145 pasiënte is 9 pasiënte uitgesluit as gevolg van die uitsluitingskriteria en daar was 2 pasiënte wat versuim het om op te volg. Twee-en-veertig pasiënte (met geen bekende vorige mediese probleme nie) het komplikasies gedurende hulle voorgeboorte verloop ontwikkel wat gelei het tot verandering in kliniese hantering en vroeëre verlossing (≤39 weke) genoodsaak het. Van die oorblywende 92 pasiënte in die oudit, het 47 (51%) in spontane kraam gegaan voor hulle induksiedatum. Daar was geen intra-uteriene sterftes voor verlossing nie. Gevolgtrekkings Noukeurige opvolg by ‘n hoërisiko kliniek identifiseer nuwe of versteekte moederlike en fetale komplikasies in 29% van pasiënte met ‘n vorige intra-uteriene sterfte sonder enige duidelike moederlike of fetale siekte in die indeks swangerskap. Wanneer alle risikos uitgesluit word en die swangerskap toegelaat word om voort te gaan tot 39 weke voor ‘n induksie aangebied word, sal 51% van pasiënte spontaan in kraam gaan.

Thesis (D.Phil.)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: Chapter 1 provides literature based background information on the clinical importance of sperm morphology as recorded by strict criteria during the diagnostic approach of the infertile couple. Furthermore, the use of a sequential diagnostic schedule for couples in an assisted reproductive programme is emphasized. The author revisited the literature on chromatin packaging of spermatozoa and addresses this issue as an additional semen parameter providing information relating to DNA damaged spermatozoa. The chapter also includes evidence underlining the growing need for the implementation of the acrosome reaction as an important contribution to the assisted reproductive programme. Chapter 2 provides detailed descriptions of the material and methods used during the study. Chapter 3 is sub-divided into 5 sections, each of which represents a separate study that was prepared as a scientific paper. The study included 338 couples consulting for infertility treatment at various gynaecologists in Pretoria and Johannesburg. The diagnostic assisted reproductive laboratory support was provided by the Andrology laboratory of Drs du Buisson and partners from Pretoria. In the first study the role of chromatin packaging as an indicator of in vitro fertilization rates, the semen samples from 72 men were used to record their chromatin packaging quality as well as their sperm morphology classification. Significant different percentages CMA3staining (mean±SE) were recorded among the 2 morphology groups, namely 65.9%±3.5 and 44.5%±1.7 (p=0.001). Using cut off values for chromatin packaging established during the first study, the second study utilized semen from 140 men in the in vitro fertilization (IVF) and intracytoplasmic sperm injection programme (ICSI) to analyze for sperm concentration, motility, morphology and chromatin packaging (CMA3).IVF and ICSI data were stratified using 3 basic cut off values for CMA3staining, namely <44%, >44-60% and >60%. The study concluded that results on the chromatin packaging quality of spermatozoa could be used as an additional parameter of sperm quality since it could provide valuable information on decondensation status of a given sperm population. The third study aimed to establish zona pellucida induced acrosome reaction response (ZIAR) among 35 couples with normal and G-pattern sperm morphology and repeated poor fertilization results during assisted reproduction treatment. Interactive dot diagrams, divided patients into 2 groups i.e. ZIAR<15% and ZIAR>15% with mean fertilization rates of 49% and 79%, respectively. The study concluded that the ZIAR test has diagnostic potential, since it can assist the clinician to identify couples that will benefit from ICSI therapy. The forth study revisited the importance of micro-assay for acrosome reaction determinations in a diagnostic andrology laboratory. The micro-assay not only allows the use of a single zona pellucida, but also facilitates the future possibility of using recombinant zona pellucida proteins in a diagnostic test system. The final study in Chapter 3 includes results obtained from 49 couples (172 oocytes) and aimed to evaluate the role of chromatin packaging and sperm morphology during sperm-zona binding, sperm decondensation and the presence of polar bodies among 170 oocytes that failed in vitro fertilization (IVF). Odds ratio analyses indicated that being in the a group with elevated CMA3 staining i.e. >60%, the risk of decondensation failure increases 15.6 fold relative to normal CMA3 staining <44%. Chapter 4 underlines the validity of the sequential diagnostic approach and summarizes the results and value of a multistep diagnostic scheme. The chapter concludes with the recommendation that both chromatin packaging quality and zona pellucida mediation of the acrosome reaction should be part of the diagnostic tools in the assisted reproductive programme. / AFRIKAANSE OPSOMMING: Die literatuuroorsig in Hoofstuk 1 konsentreer in hoofsaak op die kliniese belang van sperm morfologie en die uitbreiding van die diagnostiese toetse en hantering van die egpaar in die reproduktiewe ondersteuningsprogram. Die kromatien pakkingskwaliteit van die spermsel word onderskryf as In belangrike toevoeging tot die diganostiese program, aangesien ONS skade dikwels saam met kromatiendefekte aangetref word. Die rol van die akrosoomreaksie word ook in detail literatuuroorsigtelik beklemtoon. Hoofstuk 2 bevat volledige inligting omtrent materiaal en metodes wat in die studie gebruik is. Hoofstuk 3 bevat die eksperimentele gegewens wat in 5 afsonderlike sub-afdelings as wetenskaplike publikasies aangebied word. Die studies bestaan uit data van 338 pasiënte, wat deur verskillende ginekoloë van Pretoria en Johannesburg gekonsulteer is waartydens drs. du Boisson en vennote van Pretoria die diagnostiese reproduktiewe laboratoriumdienste verskaf het. Die eerste studie stel dit ten doel om die belang en korrelasie van die spermsel kromatienpakkingskwaliteit van 72 mans te vergelyk met die morfologiese bou van sie sel. Aangesien morfologie reeds gevertig is as 'n kliniese voorspeller van bevrugting was dit nodig om hierdie parameter te vergelyk met die kromatienpakking van die sel. Twee afsnypunte word vir die normo-en teratozoospermiese mans identifiseer naamlik, 44.5%±1.7 en 65.9%±3.5 (p=O.001),respektiewelik. Die tweede studie gebruik die afsnypunte 44% en 66% om die in vitro bevrugting en intrasellulêre sperm inspuiting (ICSI) data te ontleed. Die resultate dui aan dat kromatienpakking In waardevolle bydrae tot die diagnostiek van die pasiënte lewer. Die derde studie stel dit ten doelom die waarde van die zona pellucida geinduseerde akrosoomreaksie (ZIAR) te bepaal. Die studie sluit die data van 35 egpare in wat almal normale of G-patroon morfologie het en verder onverklaarde swak bevrugtings resultate tydens in vitro bevrugtingsterapie. Interaktiewe punt diagram (interactive dot diagrams) verdeel die data in twee groepe naamlik, ZIAR<15% en ZIAR>15% met gemiddelde bevrugtingssyfers van 49% en 79%, respektiewelik. Die studie sluit af met die gedagte dat die ZIAR toets 'n groep pasiënte identifiseer met 'n besondere fisiologiese afwyking d.i. subnormale akrosoom respons op zona pellucida blootstelling. Die vierde afdeling van die hoofstuk onderstreep die belang van die mikro-tegniek vir die bepaling van die akrosoom reaksie, wat tydens die projek gebruik is Die vyfde afdeling van Hoofstuk 3 stel dit ten doelom 170 onbevrugde eierselle van 49 pasiënte te ontleed vir moontlike oorsake vir die mislukte bevrugting. Ondersoeke sluit in die kromatienpakking, sperm-zona binding, sperm dekondensasie en die teenwoordigheid van polêre liggaampies. Statisties blyk dit dat indien 'n kromtienpakking nie normaal is nie (>66%) het die spermsel 'n 15 keer groter kans om nie te dekondenseer nie. Hoofstuk 4 bespreek die noodsaaklikheid van die diagnostiese skedule by die hantering van die onvrugbare egpaar in.

Thesis (DSc)--Stellenbosch University, 2012. / Please refer to full text for abstract.

Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: CHAPTER 1 In this chapter the aim is to outline the different chapters under section A. Against this background, we will conduct a literature review of relevant studies performed, and evaluate their comments regarding identifying embryo markers which can be utilized to improve overall ART outcome. We will evaluate the embryo marker sHLA-G in detail, using a prospective randomized study as well as a retrospective multi-centre study. The role of the morphology and genetic profile of an oocyte, zygote and embryo and subsequent blastocyst formation will be evaluated with the help of WGA/CGH. The work will then be summarized and conclusions will be made as well as possible suggestions for future directions will be indicated. In section B the methodology of the studies explaining the role of the candidate is illustrated. CHAPTER 2 In this chapter the impact of the oocyte/zygote and the embryo on implantation/pregnancy rate was discussed. The morphologic characteristics of the oocyte, the cumulus–oocyte-complex (COC), the zona pellucida, the perivitelline space, cytoplasm and meiotic spindle and the polar body and its appearance were discussed in detail. The morphologic characteristics of embryo fragmentation and its effect on embryo development, ploidy and blastocyst formation were also studied. Embryo markers to predict pregnancy outcome were researched based on the international literature. The pronuclear morphology and early cleavage were highlighted as non-invasive embryo markers to predict outcome. A non-invasive biochemical marker, soluble Human Leucocyte-Antigen-G (sHLA-G), that is expressed by developing embryos was researched. The value of blastocyst transfer and the improved ongoing pregnancy rate compared to cleavage stage embryos were highlighted based on a recent meta-analysis. A detailed discussion on sHLA-G as well as Array-CGH and the future of these tests followed. CHAPTER 3 In this chapter the aim was to compare pregnancy and implantation rates when embryos are selected based on a single Day 3 (D 3) morphology score vs. a GES score plus sHLA-G expression. This was a prospective randomized study (n=214) undergoing fresh ICSI cycles. Embryos were selected for transfer based on either Day 3 morphology score (Group A) or GES-scoring plus sHLA-G expression (Group B). The following results were reported: Clinical [35/107 (33%) vs. 52/107 (49%)] and ongoing pregnancy [20/107 (19%) vs. 52/107 (49%)] rates were significantly different between Group A and Group B (p<0.05). Implantation rates were not significantly different between Group A [52/353 (15%)] and Group B [73/417 (18%)] (p<0.05). The number of pregnancies lost during the first trimester was nearly 12 times higher in Group A [25/52 (48%)]. It was concluded that the miscarriage rate was significantly lower in Group B than Group A and the pregnancy results were superior when embryos were selected based on GES plus sHLA-G expression. CHAPTER 4 Several studies have reported an association between the presence of soluble human leukocyte antigen-G (sHLA-G) in human embryo culture supernatants (ES) with implantation and pregnancy outcome in vitro. However, the actual presence role during implantation and effect on implantation and pregnancy outcome are still controversial. A retrospective multi-centre study was performed on 2040 ICSI patients in six different centers. All embryos were individually cultured and a chemiluminescence enzyme-linked immunosorbent assay (ELISA) was used to detect the presence of sHLA-G in culture medium surrounding embryos. In all centers, a positive sHLA-G result was associated with an increase in odds of multiple clinical implantations (OR: 1.48, 95% CI: 1.07 to 2.05, p-value: 0.0170), and an increased odds of multiple on-going pregnancies (OR: 1.66, 95% CI: 1.10, 2.51, p-value: 0.0170). Data from this multi-centre study emphasize that sHLA-G expression is a valuable non-invasive embryo marker to assist in improving pregnancy outcome with the theoretical potential to reduce multiple pregnancies. A combination of sHLA-G expression and extended embryo culture to the blastocyst stage might provide future tools by which to select single embryos for transfer and reduce the risk of multiple gestational, without compromising their pregnancy rates. CHAPTER 5 In this chapter the ploidy status of first and second polar bodies and Day 3 blastomere, embryo morphology and biochemical (sHLA-G) characteristics were correlated with blastocyst development and subsequent pregnancy outcome. All oocytes/zygotes and embryos were individually cultured to the blastocyst stage. PB-I, PB-II and blastomeres underwent whole genome amplification (WGA) and comparative genome hybridization (CGH) and complete karyotyping. Each embryo‟s culture medium supernatant was collected and analyzed for sHLA-G expression on Day 2. The following results were reported: Fifty seven mature (MII) donor oocytes were obtained, 33/57 (57.9%) were aneuploid, 21/57 (36.8%) were euploid and 3/57 (5%) were “inconclusive”. No correlation was found between CGH status of PB-I, PB-II and the GES-score. Furthermore, no correlation was established between PB-I CGH results and blastocyst morphology grade. There was a significant correlation between PB-I CGH and blastomere CGH results. Euploid and aneuploid PB-I developed into 58% and 67% blastocysts, respectively. Kappa statistics (>0.7) revealed a positive correlation between the ploidy of PB-I, PB-II and the blastomeres. It was concluded that following ICSI and sequential genetic karyotyping of the oocyte/zygote and subsequent blastomeres, the majority of oocytes fertilized and subsequent zygotes developed into blastocysts, despite their ploidy status. We therefore conclude that blastocyst development is not associated with ploidy. CHAPTER 6 Identifying a developmentally competent embryo to transfer that has the highest probability to develop into a live baby has been an issue of debate and continues research. The aim of this chapter is to discuss the morphological, biochemical and genetic features of an embryo that has been shown to be predictive of implantation and pregnancy outcome in ART using most current evidence. A literature search was performed looking at the correlation between pronuclear morphology, early cleavage, cleavage stage embryos, blastocyst development, the presence of sHLA-G, CGH, embryo development and implantation/pregnancy rates in ART. Based on the available literature, a combination of observations could assist the scientist with embryo selection. The pronuclear stage morphology, the early embryo division, cleavage embryo stage and quality of the day 3 embryos provides limited guidance. However, choosing a blastocyst with a positive sHLA-G result on Day 5 is the optimal combination to make the final selection before embryo transfer or freezing. This non-invasive approach should improve pregnancy outcome and reduce multiple pregnancy rates. As far as the use of the more invasive technology such as aCGH is concerned, more research on pregnancy outcome is needed. CHAPTER 7 A combination of observations for embryo selection, starting with oocyte grading, pronuclear stage morphology, early zygote cleaving and cleavage-stage embryo morphology/quality on Day-3, however, ultimately using extended embryo culture and choosing a blastocyst on Day 5 with positive sHLA-G values available, will assist the scientist in making the final decision before selecting an embryo for transfer or cryopreservation. The use of aCGH (for chromosomal analysis) is invasive and is still considered experimental. Finally we conclude that despite all the above mentioned parameters to select an embryo for transfer that will develop into a live baby, more extensive research and international corroboration is needed in order to improve and standardize embryo selection criteria. / AFRIKAANSE OPSOMMING: HOOFSTUK 1 Die doel in hierdie hoofstuk is om die verskillende hoofstukke onder Afdeling A uiteen te sit. Daar word beplan om „n literatuur oorsig te doen van toepaslike studies rakend embriomerkers wat swangerskap-uitkoms in in vitro bevrugting kan verbeter. Verder sal die embriomerker sHLA-G deeglike bestudeer word met behulp van „n prospektiewe gerandomiseerde studie, asook „n retrospektiewe multisentrum studie. Die rol van embrio morfologie en die genetiese profiel van die ovum, sigoot asook die embrio en die daaropvolgende blastosist vorming sal geëvalueer word met behulp van WGA/CGH. Alle bevindings sal daarna opgesom word, gevolg deur „n sinvolle gevolgtrekking en laastens sal voorstelle gemaak word vir toekomstige navorsing op die gebied. In Afdeling B sal die metodiek van die studies verduidelik word, asook „n beskrywing gegee word van die kandidaat se rol gedurende die navorsings projekte in hierdie tesis. HOOFSTUK 2 In hierdie hoofstuk word die impak van die oösiet en die embrio op die inplanting/swangerskap-koers bespreek. Die morfologiese eienskappe van die oösiet, die kumulus-oösiet kompleks, die sona pellucida, die perivitelline spasie, sitoplasma en meiotiese spoel, die poolliggaam en die se voorkoms word breedvoerig bespreek. Die morfologiese eienskappe van die embrio, fragmentasie en die invloed daarvan op die embrio, ploïdie, en blastosistvorming word bespreek. Embriomerkers om swangerskapsuitkoms te voorspel, gebaseer op internasionale literatuur, is ook nagevors. Die pronukleêre morfologie en vroeë deling word as nie-indringende embriomerkers uitgelig om swangerskapsuitkoms te voorspel. „n Biochemiese, nie-indringende merker wat deur ontwikkelende embrios uitgedruk word, oplosbare menslike leukosiet antigeen-G (sHLA-G), word bespreek. Die waarde van blastosist oordrag en die verbeterde koers van voortgaande swangerskappe in vergelyking met verdelende embrios, is ook uitgelig, gebaseer op „n onlangse metanalise. „n Breedvoerige bespreking van sHLA-G asook “Array-CGH” en die toekoms van hierdie toetse word behandel. HOOFSTUK 3 Die doel van hierdie hoofstuk is om swangerskap en inplantingskoerse te vergelyk wanneer embrios geselekteer word op „n enkel Dag 3 (D 3) morfologie beoordeling, teenoor „n kumulatiewe GES-telling plus sHLA-G uitdrukking. Hierdie was „n prospektiewe ewekansige studie (n=214) waar pasiënte ICSI-siklusse ondergaan het. Embrios is geselekteer vir terugplasing gebaseer op óf Dag 3 morfologie telling (Groep A), óf „n kumulatiewe GES-telling plus sHLA-G uitdrukking (Groep B). Die volgende resultate is gerapporteer: kliniese swangerskappe [35/107 (33%) vs 52/107 (49%)] en voortgaande swangerskappe [20/107 (19%) vs. 52/107 (49%)] se sukses koerse is beduidend verskillend tussen Groep A en Groep B (p<0.05). Inplantingskoerse is nie beduidend verskillend tussen Groep A [52/353 (15%)] en Groep B [73/417 (18%)] (p<0.05) nie. Die aantal swangerskappe wat tot niet gegaan het tydens die eerste trimester was bykans 12 keer hoër in Groep A [25/52 (48%)]. Die slotsom was dat die miskraamsyfer beduidend laer in Groep B as in Groep A is en die swangerskap syfer betekenisvol beter was wanneer die selektering van embrios op GES plus sHLA-G gebaseer is. HOOFSTUK 4 Verskeie studies het „n assosiasie getoon tussen die teenwoordigheid van oplosbare menslike leukosiet antigeen-G (sHLA-G) in menslike embrio kultuur en swangerskaps uitkoms in vitro. „n Retrospektiewe studie is op 2040 ICSI pasiënte by 6 verskillende sentra gedoen om die effek van s-HLAG verder te bestudeer. Alle embrios is individueel gekweek om die teenwoordigheid van sHLA-G in „n kultuurmedium rondom die embrios te identifiseer. In alle sentra is „n positiewe sHLA-G uitslag met „n toename in die waarskynlikheid van veelvuldige inplantings geassosieer (OR: 1.48, 95% CI: 1.07 tot 2.05, p-waarde: 0.0170), asook „n toename in waarskynlikheid van meervoudige swangerskappe wat voortduur (OR: 1.66, 95% CI: 1.10, 2.51, p-waarde: 0.0170). Data uit die multisentriese studie beklemtoon dat sHLA-G uitdrukking „n waardevolle nie-indringende embriomerker is om by te dra tot die verbetering van swangerskapsuitkoms, asook die teoretiese potensiaal om meervoudige swangerskappe te verminder. „n Kombinasie van sHLA-G uitdrukking en verlengde embrio kultuur tot die blastosist stadium mag moontlik „n toekomstige hulpmiddel wees waardeur enkele embrios vir terugplasing geselekteer kan word. Daardeur kan die risiko van meervoudige swangerskappe beperk word sonder om die swangerskapkoerse in gevaar te stel. HOOFSTUK 5 In dié hoofstuk word die ploïdie status van die eerste en tweede poolliggaampies en Dag 3 blastomere, embrio morfologie en biochemiese (sHLA-G) eienskappe gekorrelleer met blastosist ontwikkelling en uiteindelike swangerskapsuitkoms. Alle oösiete/sigote en embrios is individueel tot die blastosist stadium gevolg. PB-I, PB-II en blastomere het “volledige kariotipering ondergaan deur gebruik te maak van die toets “comparative genome hybridization (CGH)”. Elke embrio se kultuurmedium supernatant is versamel en ontleed vir sHLA-G uitdrukking op Dag 2. Die volgende uitslae is gerapporteer: Sewe-en-vyftig mature (MII) donor oösiete is verkry; 33/57 (57.9%) is aneuploïd, 21/57 (36.8%) is euploïd en 3/57 (5%) is onbeslis. Geen verwantskap is gevind tussen CGH status van PB-I, PB-II en die GES-telling. Geen verwantskap is gevind tussen CGH status van sHLA-G. Verder was daar geen verwantskap gevind tussen PB-I CGH uitslae en blastosist morfologie graad nie. Daar was „n beduidende korrelasie tussen PB-I CGH en blastomeer CGH uitslae. Euploïde en aneuploïde PB-I het onderskeidelik in 58% en 67% blastosiste ontwikkel. Daar is „n positiewe verwantskap tussen die ploïdie van PB-I, PB-II en die blastomere aangetoon [Kappa (>0.7)]. Dit is afgelei dat na ICSI en sekwensiële genetiese kariotipering van die oösiet/sigoot en daaropvolgende blastomere, die meerderheid oösiete bevrug is en die daaropvolgende sigote ontwikkel het tot blastosiste, ongeag hul ploïdie status. Ons afleiding is dus dat blastosist ontwikkelling nie aan ploïdie verwant is nie. HOOFSTUK 6 In hierdie hoofstuk bespreek ons waarnemings wat betref seleksie kriteria om die beste embrios te kies vir terugplasing wat uiteindelik tot „n suksesvolle swangeskap sal lei. Morfologiese, biochemiese en genetiese faktore is ondersoek. „n Onderskeiding is gemaak tussen nie-indringende (mikroskopiese en biochemiese) en indringende (embrio biopsie, aCGH) tegnieke. 'n Kombinasie van nie-indringende observasies, wat insluit pronukliere mofologie, vroee sigoot verdeling en vroeë embrio morfologie/kwalitieit op Dag-3 het beperkte inligting verskaf wat betref swangerskapkans. Verlengde embrio kweking tot die blastosist stadium (Dag-5) plus „n positiewe sHLA-G resultaat gee egter veel meer voordelige inligting aan die embrioloog met die embrio seleksie proses, voor embrio terugplasing of bevriesing. Laasgenoemde inligting sal die swangerskap syfer bevoordeel en die meervoudige swangerskap kans verlaag. Wat die indringende tegniek (aCGH) betref, word veel meer data benodig rakend die potensiele voor- en nadele wat betref swangerskap uitkoms, voordat „n sinvolle gevolgtrekking gemaak kan word. HOOFSTUK 7 „n Volledige literatuur oorsig dui daarop dat alle beskikbare riglyne om embrios te kies vir terugplasing, ingespan moet word. In die studie is daar gekyk na „n kombinasie van hierdie voorstelle. Daar is begin met die morfologie van die pronukliere stadium, gevolg deur vroeë sigoot-verdeling, asook beoordeling van embrios se morfologie/kwaliteit op Dag-3 van ontwikkeling. Daar word voorgestel dat die keuse van „n blastosist op Dag 5, gekombineerd met „n positiewe oplosbare menslike leukosiet antigeen G (shla-G) die embrioloog van hulp kan wees om die beste embrio te kies vir terugplasing of bevriesing. Hierdie nie-indringende riglyn behoort swangerskap-uitkoms te verbeter asook meervoudige swangerskappe te verminder. Indringende tegnieke soos ACGH benodig verdere in diepte navorsing en data verkryging om die waarde van hierdie toets te kan beoordeel.

Thesis (M.D.)-University of Natal, Durban, 1979.

Background: Multiple factors contribute to mother-to-child transmission (MTCT) of HIV-1, including virological, obstetric and biological factors. Other possible contributory determinants for high MTCT rates include immunological factors such as host genetics and viral genetic variations. Despite several therapeutic, prophylactic and obstetric interventions to reduce the proportion of infants infected during labour and delivery, mechanisms for intrapartum HIV-1 transmission remain elusive and current interventions, could, therefore remain sub-optimal. Much controversy has surrounded the correlation of HIV-1 RNA (viral load) in the systemic and genital compartments of women. The influence of short-term antiretroviral (ARV) drugs on genital tract HIV-1 is also unclear. At the time the present study was initiated, a regimen of maternal intrapartum and neonatal postpartum single-dose Nevirapine (sdNVP) was the standard of care for the prevention of mother-to-child transmission (PMTCT). In most low and middle-income countries, including South Africa, sdNVP has been documented as effective intrapartum HIV-1 prevention based on plasma pharmacokinetic levels, decreased viral loads (HIV-1 RNA) and reduced rates of intrapartum transmission, yet operational studies continue to report high intrapartum transmission rates despite the administration of sdNVP. As a result perinatal HIV-1 transmission remains a significant public health concern in several African countries. Aim: The primary aim of this study was to describe the pharmacological dynamics of Nevirapine in association with virological and immunological risk factors for intrapartum HIV-1 transmission in a South African PMTCT programme where sdNVP was the standard of care. Methods: Following regulatory approval from the Biomedical Research Ethics Committee at the University of KwaZulu-Natal (UKZN), one hundred and twenty pregnant HIV-infected women who received the sdNVP regimen for prevention of mother-to-child HIV-1 transmission were enrolled between April-December 2006 at King Edward VIII Hospital (KEH) in Durban. Blood and cervicovaginal lavage (CVL) samples were collected from women at pre-NVP (during pregnancy) and post-NVP dosing (during labour/delivery). In addition to infant blood sampling at birth (post-NVP), postnatal infants were assessed at four and six weeks postnatally. Pharmacological laboratory investigations involved measurement of NVP drug concentration by Tandem Mass spectrophotometry. Virological investigations comprised HIV-1 RNA (viral load) quantitation, HIV-1 drug resistance testing (HIV-1 transmitting women only) and HIV-1 DNA PCR testing (infants only). Immunological investigations were only undertaken in a selected case-control subset of HIV-1 transmitting women and their infants. In this component, laboratory investigations included the determination of CCL3 and CCL3-L1 gene copy numbers, identification of single nucleotide polymorphisms (SNP’s) and haplotype characterisation of the CCL3 gene. All women were also screened for the presence of sexually transmitted infections (STI’s) during pregnancy. Results: One hundred and twenty women were enrolled onto this study. Of these, 110 women delivered 117 live infants (103 singletons and 7 twin pairs). Twelve (10.9%) women transmitted HIV-1 to their infants, while 95 (86.0%) were classified as non-transmitters. As a result of seven twin deliveries, the infant cohort comprised of 117 infants in total. Following two separate DNA PCR tests, HIV-1 infection was identified in 14 (11.9%) of study infants while the remaining 90 (76.9%) were exposed-uninfected. HIV infection status remained unknown for 13 infants due to infant demise (1.7%), lost to follow-up (7.7%) or study withdrawal (1.7%). During active labour (sampling that was best representative of the intrapartum phase) and within 20 hours of dosing, the median NVP concentration of 1070 ng/ml in the maternal systemic compartment was almost 44 times higher than the NVP levels detected in the genital compartment [24.5 ng/ml] (p < 0.001). NVP drug levels were below the 100 ng/ml therapeutic target in seven (13.7%) of 51 plasma and in all 39 CVL samples. While no significant association was found between NVP concentration in the systemic compartment and HIV-1 transmission (p = 0.4), this association was statistically significant in the genital compartment(p = 0.02). The median plasma NVP level detected among infants at birth was 83 times above the IC50 WT (10 ng/ml) and eight times higher than the 100 ng/ml therapeutic target for NVP. More than 71.0% of the infants achieved NVP drug levels above the therapeutic target. In general, higher levels of HIV-1 RNA (viral load) were observed in maternal plasma when compared to CVL. Following intrapartum sdNVP dosing, reduction in HIV-1 RNA levels did occur, however R80.0% of the women experienced no change to their HIV-1 RNA levels in both systemic and genital compartments during active labour. These findings were further supported by the strong correlation observed when comparing pre and post-NVP HIV-1 RNA levels in both maternal systemic [r = 0.81, p < 0.0001] and genital compartments [r = 0.80, p < 0.0001] during active labour. HIV-1 transmitting women had significantly higher viral loads than their non-transmitting counterparts in systemic and genital compartments, before and after intrapartum sdNVP administration. In terms of perinatal transmission this observation was only statistically significant for plasma (p = 0.02) and not CVL (p = 0.7). Maternal viral load was inversely correlated with maternal CD4 cell counts in both systemic and genital compartments. Almost 40.0% of women in this study had at least one type of STI detected during pregnancy. Maternal STI’s were detected in four (66.6%) intrapartum transmitting women and in 38 (38.8%) of non-transmitting women. No significant association was observed between the presence of maternal STI’s and the risk for intrapartum MTCT (p = 0.2,RR: 2.90, 95% CI: 0.60-15.40). The presence of maternal STI’s was associated with higher median viral loads in both systemic and genital compartments of all women, independent of intrapartum HIV-1 transmission. Despite trial-like conditions and optimal sdNVP dosing, the overall MTCT rate in this exclusively formula-fed cohort was 11.9%, of which 50.0% were in utero and 50.0% were intrapartum HIV-1 transmissions. In utero and intrapartum MTCT rates were 5.9% and 5.9% respectively. Discussion/Conclusion: Detectable CVL HIV-1 RNA that correlated well with plasma HIV-1 RNA, in conjunction with sub-optimal NVP drug concentration in maternal CVL during active labour, suggests that intrapartum HIV-1 infected women continue to act as reservoirs for both vertical and horizontal HIV-1 transmission throughout the duration of pregnancy. These findings confirm that the role of sdNVP in PMTCT was primarily one of infant prophylaxis. This was further supported by relatively unchanged maternal HIV-1 RNA (viral load) during active labour, in both systemic and genital compartments. Early identification of women who need highly active antiretroviral therapy (HAART), and initiation of such therapy as early as possible during pregnancy, not only benefits maternal health but remains the best prophylaxis against mother-to-child HIV-1 transmission. Universal access to HAART and improving strategies to optimize coverage of the current dual ARV regimen sdNVP and Zidovudine for PMTCT remain urgent research priorities in several resource-limited settings. Ongoing STI counseling, intensive screening/testing of women and their partners together promotion of condom usage, safer sex practices and aggressive STI treatment are simple interventions with tremendous impact for PMTCT in resource-limited settings. / Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2009.

Obstetric trauma is a major cause of faecal incontinence in women resulting in considerable social disability. Until recently the development of faecal incontinence has been attributed largely to damage to the pelvic nerves. However the advent of anal endosonography has added a new dimension to our understanding of the pathophysiology of faecal incontinence. In this thesis, gross dissection and histological studies of 19 anorectal specimens was performed to demonstrate the normal anatomy. Simultaneous dissection and sonography of the anorectum (14 in vivo and 12 in vitro studies) has clarified the normal sonographic anatomy of the anal sphincters. Anal endosonography was performed in 114 healthy volunteers to demonstrate gender differences in anal sphincter anatomy. A prospective study of 12 patients undergoing secondary sphincter repair and 15 patients undergoing lateral internal anal sphincterotomy has validated the appearance of sonographic sphincter defects. A new technique of demonstrating the anal sphincters at rest using vaginal endosonography has been demonstrated in 20 women. A prospective study of 202 pregnant women using anal endosonography and neurophysiological tests has demonstrated that 35% of primigravidae (13% symptomatic) and 44% of multigravidae (23% symptomatic) develop occult anal sphincter defects during vaginal delivery. Although pudendal nerve damage can be identified in 16% of women 6 weeks after delivery, in the majority this recovers with time. Forceps delivery was identified as the single independent variable associated with sphincter damage although damage was also sustained in the absence of instrumental delivery. In a separate study of 50 women who sustained a recognized third degree tear 47% were found to be symptomatic despite a primary sphincter repair. In 85% of these women persistent anal sphincter defects were identified sonographically. In a further study of 43 women who had an instrumental delivery (17 vacuum and 26 forceps) anal sphincter defects were identified in 81% (38% symptomatic) of women who were delivered by forceps compared to 12% (21% symptomatic) delivered by the vacuum extractor. One hundred and fifty doctors and midwives were interviewed to assess their knowledge and training in perineal anatomy and repair. There was a clear deficiency in knowledge and inconsistencies in classification of third degree tears were apparent highlighting the need for more focused training in perineal anatomy and repair. / Thesis (Ph.D.)-University of Natal, 1994.