Thesis (MScMedSc (Pathology. Medical Microbiology))--University of Stellenbosch, 2005. / The aim of this study was to investigate the anti-inflammatory and immunosuppressive effects of the synthetic progestins, MPA and NET-A on human cells in vitro. These injectable contraceptives are used extensively throughout the world, including Africa. The potential of these two synthetic hormones to have certain immunosuppressive and GC properties have previously been shown. Therefore, it was of concern to us to investigate whether these two hormones could possibly demonstrate any of these GC-like properties at contraceptive doses. This was achieved by determining the effects of these two synthetic hormones in vitro on certain immunologic parameters. Chapter 1 is a literature review on MPA, NET and GCs. This chapter starts with a short introduction that sets the scene. The mode of action, effectiveness, sideeffects as well as previously reported relevant data on both MPA and NET-A is portrayed in this review. Research on the known GC, Dex, is also included in the section dealing with GCs, because this synthetic hormone was used as a comparative GC in all our experiments. This chapter soon makes the reader realize how much evidence exists that indicate the possible immunosuppressive effects these two contraceptive hormones, in particular MPA, could have. The possible anti-inflammatory or pro-inflammatory effects of MPA and NET-A are investigated in Chapter 2. This was done in vitro by measuring the effects of these two synthetic hormones on the inflammatory markers, IL-6 and TNFα, by means of ELISA. In this chapter we demonstrate that MPA, even at contraceptive doses, exhibits significant anti-inflammatory properties on both cytokines tested, while NETA displayed considerably less anti-inflammatory tendencies. In its true antiinflammatory manner, we found that Dex significantly inhibited the release of both inflammatory markers from human monocytes. In Chapter 3, we investigated the effects of MPA and NET-A on the activation of human lymphocytes. This was achieved by flow cytometric measurement of the expression of the activation membrane marker CD69 by CD4 and CD8 T cells. Here we discovered that MPA had a very significant inhibitory effect on the activation of both CD4+ and CD8+ T cells, while NET-A only significantly inhibited the activation of CD8+ T cells. In addition, we found that the inhibition of CD4+ and CD8+ T cell activation by MPA was more or less the same as the known GC, Dex, and in some cases even more potent. Chapter 4 consists of an investigation of the effects of MPA and NET-A on the cytokines belonging to TH1 and TH2 subsets of CD4 T cells. This was achieved by determining whether MPA and/or NET-A targeted specific subsets of T helper cells by measuring the distinct regulatory cytokines, IFNγ and IL-4. The mechanism and role of the T helper subsets are discussed in the introduction of this chapter. Our results were portrayed as a ratio of TH2: TH1 on which the statistical analysis was done. In addition to the analysis done on the ratio, we analyzed the helper subsets separately in order to determine which subset(s) were influenced. The results of this chapter showed that neither MPA nor NET-A significantly affected either one of the helper subsets, while Dex significantly decreased this ratio. After our observed effects of MPA and NET-A on CD8 T cells, it became of interest in Chapter 5 to investigate the effects of these two synthetic hormones on the CD8 T cell-specific chemokine, RANTES. This was achieved by measuring the effects MPA and NET-A had on RANTES production in vitro by means of ELISA. Surprisingly, we discovered in this chapter that MPA and NET-A enhanced RANTES production before and after activation of CD8 T cells. We also found that Dex had the same effect on RANTES production, but to a lesser degree. Finally, a general conclusion depicting the significance and implications of our results as well as possible future research that is required is presented in Chapter 6. It was of great importance to discuss and interpret the magnitude of data generated out of all our experiments to the utmost of our capabilities. We found that MPA, even at contraceptive doses, displayed significant immunosuppressive as well as anti-inflammatory properties. NET-A, on the other hand, demonstrated weaker immunosuppressive properties in our research and no significant anti-inflammatory properties. These findings could have clinical implications in females being treated with these synthetic contraceptives. We also demonstrated significant variation found amongst genders in response to MPA, NET-A and Dex.

Thesis (MScMedSc) -- University of Stellenbosch, 2002. / ENGLISH ABSTRACT: The gag gene of HIV-1 encodes for one of the major structural proteins, which contains several conserved cytotoxic T cell (CTL) epitopes. Gag specific CTL responses are important in controlling viral load during acute infection and asymptomatic stages of the infection. Currently, only one complete South African HIV-1 subtype C gag sequence has been published. The first aim of this study was to characterise the complete gag gene of 15 HIV-1 subtype C isolates, to be used as a set of reference sequences in the design of a South African HIV-1 subtype C vaccine. Fifteen HIV-1 subtype C isolates selected for this study, were isolated during 1998 and 1999 from the HIV-1 positive patients attending the Infectious Disease Clinic at Tygerberg Hospital. The gag gene of these isolates was amplified by PCR, cloned into mammalian expression vectors and sequenced. Restriction digest analyses as well as phylogenetic analyses were performed on the sequencing data. Previously published mutational analyses and CTL epitopes were compared to the predicted amino acid sequences of the gag clones. Sequences of 23 complete gag genes representing the 15 HIV-1 subtype C isolates as well as one complete sequence of an HIV-1 subtype B isolate were compiled. Subtyping by restriction fragment length polymorphism (RFLP) would have correctly identified 14 of the 15 subtype C isolates as subtype C and one as unidentifiable. The subtype B isolate would have also been correctly identified. Phylogenetic analyses showed that our subtype C isolates clustered with reference subtype C strains from various countries, including Botswana, India, Israel, Tanzania and Zambia. Strains from Ethiopia and Brazil formed a separate subtype C cluster. The diversity between our isolates was comparable to the diversity seen between all the HIV-1 subtype C strains. Comparisons of previously published mutational analyses and CTL epitopes to the predicted amino acid sequences of the gag clones, showed conservation in most of the clones throughout the sequence. A second aim was to establish transfection and Western Blot techniques in our laboratory for use in future studies. An in vitro transcription! translation assay was performed on the gag clones and the protein producing clones were used to transfect mammalian cells using electroporation. A Western blot was then used to screen for Gag protein expression in the transfected cell Iysates. The in vitro transcription! translation assay showed that seven of the 23 clones could produce a protein of -55 kDa in size. Four out of the seven of these clones gave a weak expression of a-55 kDa protein after transfection in a mammalian cell line. Since the completion of the experimental work of this study, other cloned HIV-1 genes have successfully been transfected into mammalian cells using the electroporation technique and the proteins produced were screened for by Western blot. To conclude with; the native form of the gag gene does not elicit strong expression of the protein, but studies have shown that expression can be improved by sequence-modification of the gag nucleotide sequence. Due to the conservation of gag, the sequence of any subtype C strain can be used for the development of a Southern African vaccine. / AFRIKAANSE OPSOMMING: Die HIV-1 gag geen kodeer vir een van die hoof strukturele proterene en bevat verskeie sitotoksiese T-limfosiet epitope. Gag spesifieke sellulere immuun respons is belangrik vir die beheer van virale lading tydens akute infeksies en tydens asimptomatiese fases van die infeksie. Tans is slegs een volledige Suid Afrikaanse HIV-1 subtipe C nuklerensuur volgorde gepubliseer. Die eerste doel van hierdie studie was om die volledige gag geen van 15 HIV-1 subtipe C isolate te karakteriseer, om gebruik te word as In stel verwysings nukleiensuur volgordes, vir die ontwerp van In Suid Afrikaanse HIV-1 subtipe C entstof. Die 15 HIV-1 subtipe C isolate wat vir hierdie studie geselekteer is, is tydens 1998 en 1999 ge·lsoleer vanaf HIV-1 positiewe pasiente wat die Infeksiesiekte Kliniek, Tygerberg Hospitaal bygewoon het. Die gag geen van hierdie isolate is geamplifiseer deur PKR, gekloneer in soogdier ekspressie vektore en die nukleiensuur volgorde is bepaal. Die nuklerensuur volgorde is gebruik in restriksie ensiem analises asook filogenetiese analises. Reeds gepubliseerde mutasie analises en limfosiet epitope is met die voorspelde aminosuur volgorde van die gag klone vergelyk. Die nukleiensuur volgordes van die 23 volledige gag gene wat die 15 HIV-1 subtipe C isolate verteenwoordig, asook een volledige nukleiensuur volgorde van een HIV-1 subtipe B isolaat, is saamgestel. Subtipering deur middel van restriksie fragment lengte polimorfisme (RFLP) sou 14 uit die 15 subtipe C isolate korrek qerdentifiseer het, maar sou een nie kon identifiseer nie. RFLP sou ook die subtipe B isolaat korrek qerdentifiseer het. Filogenetiese analises het gewys dat ons subtipe C isolate met die verwysings subtipe C stamme van verskeie lande, insluitend Botswana, lndie, Israel, Tanzania en Zambie groepeer. Stamme van Ethiopie en Brasilie het In aparte subtipe C groep gevorm. Die diversiteit tussen ons isolate was vergelykbaar met die diversiteit tussen al die subtipe C stamme. Vergelykings van gepubliseerde mutasie analises en limfosiet epitope met die voorspelde aminosuur volgorde van die gag klone, het konservasie in meeste van die klone, deur die hele nukleiensuur volgorde, getoon. Die tweede doel was om die metodes van transfeksie en Westerse klad in ons laboratorium tot stand te bring. In vitro transkripsie/ translasie toetse is gedoen op die gag klone en die proteten produserende klone is gebruik om soogdierselle te transfekteer deur gebruik te maak van elektroporasie. In Westerse klad is toe gebruik om vir Gag proterenuitdrukkinq in die sellisate te toets. Die in vitro transkripsie/ translasie toets het getoon dat sewe uit 23 klone, In proteren van -55 kDa kon produseer. Vier uit die sewe van hierdie klone het In -55 kDa proteren swak uitgedruk na transfektering van soogdier selle. Sedert die voltooiing van die eksperimentele werk van hierdie stud ie, is ander gekloneerde HIV-1 gene suksesvol in soogdierselle getransfekteer met die gebruik van elektroporasie en die proterene is met In Westerse klad aangetoon. Ten slotte: die natuurlike vorm van die gag geen ontlok nie In sterk ekspressie van die proteren nie, maar ander studies het wei aangetoon dat die ekspressie verbeter kan word met modifikasie van die gag nukleiensuur volgorde. As gevolg van die konservasie van gag, kan die nuklerensuur volgorde van enige subtipe C stam gebruik word vir die ontwikkeling van In Suider Afrikaanse entstof. / The Poliomyelitis Research Foundation / The South African AIDS Vaccine Initiative / Harry Crossley Foundation

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: Introduction Worldwide, up to 25% of women are assaulted during pregnancy, with estimates varying between populations. Violence has been associated with adverse pregnancy outcome, including preterm birth, abruptio placentae and low birth weight. Among the Coloured population of the Western Cape the incidence of spontaneous preterm birth is 20%, compared to the global figure of 10%. Overall, the rate of preterm labour has not dropped over the past 40 years and no clearer answer as to a specific cause has been found. The objective of this study was to determine whether patients who deliver preterm experience more domestic violence than those who deliver at term. Methods Two groups of patients were assessed. Firstly, patients who spontaneously delivered between 24 and 33 weeks (24wOd - 33w6d), who were admitted for suppression of active labour after 24 weeks, or who experienced placental abruption before 34 weeks, were screened for domestic violence using the "Abuse Assessment Screen". A second group of women, attending a local Midwife Obstetric Unit with uncomplicated pregnancies, completed the same questionnaire. The questionnaires were all administered by the same person (J.S.) after written informed consent was given. Results A total of 229 patients were interviewed, 99 in the low risk (LR) and 130 in the preterm labour (PTL) group, which included 23 women with abruptio placentae. The PTL group experienced significantly more violence throughout their lives than the LR group (59.7% vs. 40.4%, p = 0.038). Experiences of violence within the last year or during the pregnancy did not reach statistical significance between the two groups, although the numbers were higher for the PTL group. The PTL group smoked significantly more cigarettes per day (p = 0.009), used more alcohol (p < 0.001) and had a higher incidence of syphilis than the LR group (p = 0.005). These differences remained the same when the abruptio's were analyzed as a separate group. Conclusions: Women who delivered preterm did experience more violence at some point in their lives and were also more likely to engage in high-risk behaviour. Violence alone does not seem to cause PTL directly, but is part of a low socioeconomic lifestyle. The fact that the alcohol use is so high among these women is a problem that needs to be addressed, but once again, it is possibly the result of deeper social problems. The need for education on values and respect, family planning use and low risk sexual behaviour is once again challenged. / AFRIKAANSE OPSOMMING: GEWELD TEEN VROUE -IMPAK OP REPRODUKTIEWE GESONDHEID EN UITKOMS VAN SWANGERSKAP Inleiding Daar word beraam dat tot 25% van alle swanger vroue aangerand word, maar die insidensie wissel tussen verskillende populasies. Ervarings van geweld kan 'n direkte of indirekte oorsaak wees van swak verloskundige uitkoms wat voortydse kraam, abruptio placentae en lae geboortegewig insluit. In die Wes- Kaap, onder die Kleurlingbevolking, is die insidensie van voortydse kraam 20%, wat swak vergelyk met die wêreldwye insidensie van 10%. Gedurende die laaste 40 jaar het die voorkoms van voortydse kraam nie verminder nie en geen deurbrake is gemaak t.o.v die oorsaak van die probleem nie. Die doel van hierdie studie was om te bepaal of vroue wat prematuur verlos moontlik meer geweld ervaar as vroue wat op normale swangerskapsduur verlos. Metodes Twee groepe vroue is bestudeer. Die eerste groep het vroue ingesluit wat spontaan verlos het tussen 24 en 33 weke (24wOd - 33w6d) of vroue wat na 24 weke swangerskapsduur toegelaat is vir onderdrukking van kraam. Vroue met plasentale loslating (abruptio placentae) voor 34 weke, sonder onderliggende hipertensiewe toestande, was ook ingesluit in die groep. Daar is m.b.v. 'n vraelys ("Abuse Assessment Screen") bepaal watter van die vroue gesinsgeweld ervaar het. Die tweede groep het vroue ingesluit met ongekompliseerde swangerskappe en wat by 'n nabygeleë kliniek voorgeboortesorg ontvang het. Hulle is ook gevra om die vraelys te voltooi en is opgevolg om die uitkoms van hulle swangerskappe te noteer. Die vraelyste is almal deur een persoon (J.S.) aan die vroue voorgelê nadat hulle ingeligte, skriftelike toestemming gegee het. Resultate 'n Totaal van 229 vroue was ingesluit, 99 in die lae risiko (LR) groep en 130 in die voortydse kraam (VK) groep, waarvan 23 abruptio placentae gehad het. In vergelyking met die LR groep, het die VK groep het betekenisvol meer geweld in hulle leeftyd ervaar (59.7% teenoor 40.4%, p = 0.038). Geweld wat tydens die afgelope jaar of tydens die swangerskap ervaar is, het nie betekenisvol verskil tussen die twee groepe nie, alhoewel die getalle hoër was vir die VK groep. Die VK groep het betekenisvol meer sigarette per dag gerook (p = 0.009), meer alkohol gebruik (p < 0.001) en het 'n hoër insidensie van sifilis gehad as die LR groep (p = 0.005). Hierdie verskille was steeds beduidend nadat dié met abruptio placentae as 'n aparte groep geanaliseer is. Gevolgtrekking Die vroue wat prematuur verlos het, het meer emosionele en fisiese geweld in hulle leeftyd ervaar en is meer geneig om 'n ongesonde leefstyl te handhaaf. Geweld blyk nie 'n direkte oorsaak van voortydse kraam te wees nie, maar gaan gepaard met 'n lae sosio-ekonomiese lewensstyl. Die hoë insidensie van alkoholgebruik onder swanger vroue is 'n probleem wat aangespreek moet word, maar dit is waarskynlik die manifestasie van dieper emosionele probleme. Opvoeding in terme van waardes en respek, gesinsbeplanning en veilige seksuele gedrag is gevolglik 'n noodsaaklikheid.

Thesis (MScMedSc)-- Stellenbosch University, 2013. / ENGLISH ABSTRACT: Preconceptual sex selection is an ethically justifiable process whereby X- and Y-chromosome bearing spermatozoa are isolated prior to fertilization of the oocyte in order to generate either a male or a female offspring. Although various separation techniques are available, none can guarantee 100% accuracy. There are various physiological differences between X- and Y-chromosome bearing spermatozoa which can be used to separate these two populations of sperm. For the purpose of this study, X- and Y-chromosome bearing spermatozoa were separated based on (1) their respective abilities to remain viable when subjected to adverse environments, including extreme pH values, increased temperatures and various hydrogen peroxide (H2O2) concentrations; (2) the ability of Y-chromosome bearing spermatozoa to swim faster and/or more progressively than X-chromosome bearing spermatozoa; and (3) the X-chromosome bearing spermatozoa’s increased size and weight when compared to the Y-chromosome bearing spermatozoa. The efficacy of live and dead cell separation through (i) Magnetic Antibody Cell Separation (MACS) and (ii) a modified swim-up technique was also assessed and compared. Changes in the sex-chromosome ratio of samples were established by double-label fluorescent in situ hybridization (FISH) before and after processing. Sperm motility (CASA) and viability (eosin/nigrosin) was assessed before and after each intervention. Ethical clearance for this study was granted by the Health Research Ethics Committee 1 (Ethics #: S13/04/068). The results indicated successful enrichment of X-chromosome bearing spermatozoa upon incubation in acidic media, increased temperatures, and H2O2. In contrast, Y-chromosome bearing spermatozoa were successfully enriched through a direct swim-up method as well as discontinuous gradient centrifugation. In conclusion, this study demonstrated the potential role for physiological differences between X- and Y-chromosome bearing spermatozoa in the development of preconceptual gender selection through sperm sorting. / AFRIKAANSE OPSOMMING: Prekonsepsie geslagselektering is 'n eties regverdigbare proses waardeur X- en Y- chromosoom draende spermatosoë geïsoleer word voordat bevrugting van die oösiet plaasvind, om óf 'n manlike óf 'n vroulike nageslag te genereer. Alhoewel verskeie skeidingstegnieke beskikbaar is, kan geeneen 100% akkuraatheid waarborg nie. Daar bestaan verskeie fisiologiese verskille tussen X- en Y- chromosoom draende spermatosoë wat skeiding van hierdie twee groepe spermatosoë moontlik kan maak. Vir die doel van hierdie studie is skeidingsmetodes vir die X- en Y- chromosoom draede spermatosoë gebaseer op (1) hul onderskeie vermoëns om lewensvatbaar te bly tydens blootstelling aan ‘n ongunstige milieu, insluitend ekstreme pH waardes, verhoogde temperature en verskeie waterstofperoksied (H2O2) konsentrasies; (2) die vermoë van die Y-chromosoom draende spermatosoon om vinniger en/of meer progressief as X-chromosoom draende spermatosoë te swem; en (3 ) die X-chromosoom draende spermatosoon se verhoogde grootte en gewig in vergelyking met die Y- chromosoom draende spermatosoon. Die effektiwiteit van die (i) Magnetiese Anti-liggaam Sel Skeidingstegniek (MACS) en (ii) 'n aangepaste weergawe van die op-swem tegniek om lewendige en dooie selle te skei is ook bepaal en vergelyk. Veranderinge in die geslagschromosoom verhouding van die monsters is bepaal deur dubbel-etiket fluoresensie in situ hibridisering (FISH) voor en na verwerking. Spermmotiliteit (CASA) en lewensvatbaarheid (eosien/nigrosin) is bepaal voor en na elke intervensie. Etiese goedkeuring vir hierdie studie is verleen deur die Gesondheids-Navorsingsetiekkomitee 1 (Etiese # : S13/04/068). Die resultate dui suksesvolle verryking van X-chromosoom draende spermatosoë deur inkubasie in suur media, verhoogde temperature, en H2O2. Y-chromosoom draende spermatosoë is verryk deur middel van 'n direkte op-swem metode sowel as diskontinue gradiënt sentrifugering . Ten slotte, hierdie studie toon die potensiële rol vir fisiologiese verskille tussen X- en Y- chromosoom draende spermatosoë in die ontwikkeling van prekonsepsie geslagselektering metodes deur skeiding van X- en Y-chromosoom draende sperme.

Thesis (MCur)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: The critical care nurse works in an environment where patient need often shifts the parameters within which she or he practices. It is expected of a skilled critical care nurse to be able to make independent decisions and take action regarding patient care based on her or his knowledge and skills without discounting the parameters of her or his scope of practice. Practice experience has indicated that the critical care nurse is often uncertain about whether her or his clinical activities are protected by the regulations provided by the Nursing Council. This is more specifically true in the private hospital industry where medical advice or assistance is not always easily available. This situation led to the following research question: Do the available professional and legal guidelines provide an appropriate foundation to guide the practice of the registered critical care nurse in the private hospital sector critical care environment? A non-experimental descriptive study with a qualitative orientation was conducted in 19 private hospitals in the Western Cape. Through nonprobability, random sampling, 71 registered critical care nurses were included in the study. A questionnaire was designed and validated to collect the data. Quantitative data was analysed through Excel® while qualitative data was analysed thematically. It was found that the legal and professional guidelines in place at present do provide a foundation for the clinical activities of critical care nursing in the private hospital sector. It is suggested that it is rather the critical care nurses’ interpretation of the Scope of Practice (No.R.2598 of 30/11/1984 as amended) that limits their practice as opposed to the wording of the regulations. It is recommended that critical care nurses must determine nursing care parameters based on patient need, using the regulations as a foundation for critical, analytical and reflective practice rather than as a set of rules to be followed. Key words: Scope of practice, critical care practice, ICU nursing care, private hospital nursing practice. / AFRIKAANSE OPSOMMING: Die kritiekesorgverpleegkundige werk in ‘n omgewing waar pasiëntebehoeftes gereeld die parameters waarin sy of hy praktiseer, verskuif. Dit word van ’n bekwame kritiekesorgverpleegkundige verwag dat sy of hy onafhanklike besluite en aksies met betrekking tot pasiëntesorg, gebaseer op haar of sy kennis en vaardighede, sal neem sonder om die parameters van haar of sy bestek van praktyk te oorskry. Praktykondervinding het getoon dat die kritiekesorgverpleegkundige dikwels onseker is oor watter van haar of sy optredes deur die Regulasies, soos deur die Raad op Verpleging gespesifiseer word, beskerm word. Dit is nog meer spesifiek van toepassing in die privaathospitaal-industrie waar geneeskundige advies en bystand nie altyd maklik beskikbaar is nie. Die situasie het tot die volgende navorsingsvraag aanleiding gegee: Voorsien die beskikbare professionele en wetlike riglyne ’n geskikte grondslag om die praktyk van ’n geregistreerde kritiekesorgverpleegkundige in die privaatsektor- kritiekesorgomgewing te rig? ’n Nie-eksperimentele, beskrywende studie met ’n kwalitatiewe oriëntasie is in 19 hospitale in die Wes-Kaap onderneem. Deur nie-waarskynlikheids-, toevallige steekproefneming is 71 geregistreerde kritiekesorgverpleegkundiges in die studie ingesluit. ’n Vraelys is ontwerp en gevalideer om inligting in te samel. Kwantitatiewe data is deur middel van Excel ontleed terwyl kwalitatiewe data tematies ontleed is. Daar is gevind dat die wetlike en professionele riglyne wat tans beskikbaar is, ‘n grondslag bied vir die kliniese aktiwiteite van kritiekesorgverpleegkundiges in die privaathospitaal.. Dit word voorgestel dat dit die kritiekesorgverpleegkundige se interpretasie van die Bestek van Praktyk (No.R.2598 of 30/11/1984 soos aangepas) is wat hulle praktyk beperk, eerder as die bewoording van die regulasie self.

Thesis (MScMedSc (Biomedical Sciences. Molecular Biology and Human Genetics))--University of Stellenbosch, 2009. / Hypertrophic cardiomyopathy (HCM), an inherited primary cardiac disorder mostly caused by defective sarcomeric proteins, is considered a model for studying left ventricular hypertrophy (LVH) in the absence of increased external loading conditions. The disease manifests extreme variability in the degree and pattern of LVH, even in HCM patients with the same causal mutation. The clinical phenotype of HCM can therefore be viewed as a product of the effect of sarcomere dysfunction and of additional genetic modifiers. Components of the renin-angiotensin-aldosterone system (RAAS) are plausible candidate modifiers because of their effect on blood pressure and their direct hypertrophic effect on cardiomyocytes. The present study investigated genes encoding components of the RAAS for association with cardiac hypertrophy traits, in 353 individuals comprised of genetically and echocardiographically affected and unaffected family members, belonging to 22 HCM families with HCM founder mutations by employing a multi-SNP approach with TaqMan allelic discrimination technology. Gene-gene interaction analysis was also performed to investigate the effect of epistasis on hypertrophy. Candidate genes for analysis included the angiotensin II type 2 receptor (AT2 receptor), renin, renin-binding protein (RnBP), the (pro)renin receptor, the mineralocorticoid receptor as well as genes encoding subunits of the epithelial sodium channels (ENaC) and Na+/K+-ATPase that showed evidence for cardiac expression. The present study demonstrates for the first time that variations in the renin and RnBP genes play a role in modulating hypertrophy in HCM, independent of blood pressure and confirms the involvement of the AT2 receptor in hypertrophy in HCM. Additionally we report an association between Na+/K+-ATPase α1- and β1-subunits as well as the ENaC α- and β-subunits and hypertrophy. Significant evidence for epistasis was found between renin and downstream RAAS effectors, suggesting a complex interplay between these RAAS variants and the hypertrophic phenotype in HCM. The identification of such modifiers for HCM may offer novel targets for hypertrophy research and ultimately antihypertrophic therapy.

Thesis (MScMedSc (Biomedical Sciences. Molecular Biology and Human Genetics. Medical Biochemistry))--University of Stellenbosch, 2009. / The mammalian neocortex contributes to the increasing functional complexity of the mammalian brain, partly because of its striking organisation into distinct neuronal layers. The development of the neocortex has been well studied because disrupted neurodevelopment results in several human diseases. The basic principles of neocortical development have been well established for some time; however the molecular mechanisms have only recently been identified. One major advance in our understanding of these molecular mechanisms was the discovery of Reelin, an extracellular matrix protein that directs the migration of neurons to their final positions in the developing neocortex. Reelin is a large multi-domain protein that exerts its functions by binding to its ligands on the cell surface and initiating a signal transduction cascade that ultimately results in cytoskeletal rearrangements. Several investigations have been undertaken to elucidate the functions of each of these domains to gain a better understanding reelin’s functions. We have previously identified the WR40 repeat protein 47 (WDR47), a protein of unknown function, as a novel putative ligand for the N-terminal reeler domain of reelin. To gain better understanding into the functional significance of this interaction, the present study sought to identify novel WDR47- interacting proteins. In order to achieve this, a cDNA encoding a polypeptide that contains the two N-terminal domains of WDR47, i.e. the Lis homology and the C-terminal Lis homology domain (CTLH) was used as bait in a Y2H screen of a foetal brain cDNA library. Putative WDR47 ligands were subsequently verified using 3D in vivo co-localisation. Results of these analyses showed that SCG10, a microtubule destabilizing protein belonging to the stathmin family of proteins, interacted with the N-terminal of WDR47. The identification of SCG10 as a novel WDR47 interacting protein not only sheds some light on the role and function of WDR47 but also aids in a better understanding of the reelin pathway and cortical lamination. Moreover, the data presented here, may also provide researchers with new avenues of research into molecular mechanisms involved in neuronal migration disorders.

Thesis (MScMedSc (Obstetrics and Gynaecology))--University of Stellenbosch, 2005. / CHAPTER 1 presents comprehensive background information regarding all aspects addressed in this thesis. Special attention was given to literature on paternal influences on embryonic development, the role of sperm RNA, sperm chromatin and sperm functional aspects i.e. morphology and acrosomal status and size. The experimental design and all relevant methods used during the study as well as the material that were used are presented in CHAPTER 2. The results of the different techniques and evaluations are provided in CHAPTER 3. It was found that 70% of the embryos that showed no developmental potential were Y-chromosome bearing embryos. The sperm selection process for ICSI based on the approach of choosing the “best looking“ spermatozoon in the ejaculate seem to provide cells that can be classified as normal based on the length width ratio set by the WHO for normal cells. The chromatin packaging quality of the sperm correlated significantly and negatively with the percentage normal cells in the ejaculates. CHAPTER 4 comprises of a general discussion of the results and short summary of the major findings during the project. The discussion section focused on the paternal influence on the embryonic development and provided a suggestion for future research that can possibly lead to the use of X-chromosome bearing sperm in case of severe male factor cases. CHAPTER 5 contains the bibliographical information of the study.

Thesis (MMed)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: The aim of the study was to conduct a structured review of the literature published on the use of normal sperm morphology, as an indicator of male fertility potential in intrauterine insemination (M) programs. Published literature in which normal sperm morphology was used to predict pregnancy outcome in lUI during the period 1984 - 1998 was reviewed. Four hundred and twenty one articles were identified. Eighteen provided data that could be tabulated and analyzed. Eight of the analyzed studies provided sufficient data for statistical analysis. Six studies used the Tygerberg strict criteria and two the WHO guidelines (1987, 1992). A meta-analysis of the six studies in the strict morphology group yielded a risk difference (RD) between the pregnancy rates achieved in the patients below and above the 4% strict criteria threshold of -0.07 (95% CI: -0.11 to -0.03; p< 0.001). WHO criteria group (1987,1992) had insufficient data to be analysed. Meta-analysis showed a significant improvement in pregnancy rate above 4% threshold for strict criteria. Accurate evaluation of normal sperm morphology results should be an integral part of evaluating the male factor. / AFRIKAANSE OPSOMMING: Die doel van die studie was om 'n gestruktureerde literatuuroorsig van die gepubliseerde data oor normale sperm morfologie uit te voer om vas te stelof dit enige waarde het as voorspeller van manlike fertiliteitspotensiaal in intra uteriene inseminasie (lUI) programme. Gepubliseerde literatuur waar normale sperm morfologie gebruik IS om swangerskapsuitkoms te voorspel met IUI in die tydperk 1984 - 1998 is nagegaan. Vierhonderd een en twintig artikels is geïdentifiseer. Agtien het genoeg data gehad om te kan tabuleer en analiseer. Agt van die geanaliseerde studies het voldoende data gehad vir statistiese analise. Ses studies het die Tygerberg streng kriteria gebruik en twee die WGO (1987, 1992) riglyne. 'n Meta-analise van die ses studies in die streng kriteria groep het 'n risiko verskil tussen swangerskapstempo in pasiënte onder en bo die 4% streng kriteria afsnypunt, van -0,07 (95% betroubaarheidsindeks: -0.11 tot -0.03; p<O.OOl) getoon. Die WGO kriteria (1987,1992) groep het onvoldoende data gehad om te kan analiseer. Meta-analise het 'n bekenisvolle verbetering in swangerskapuitkoms bo die 4% afsnypunt getoon vir die streng kriteria. Akkurate evaluasie van normale sperm morfologie resultate behoort 'n integrale deel te wees van die proses om die manlike faktor in infertilteitsbehandeling volledig te evalueer.

Thesis (MMed)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: OBJECTIVE: The aim of this study was to compare the accuracy of non-invasive brachial blood pressure measurements, using automated and manual devices, to invasive brachial intra-arterial blood pressure measurements in patients with preeclampsia, during acute severe hypertension. STUDY DESIGN: A prospective descriptive cross sectional study was conducted in the Obstetrics Critical Care Unit (OCCU) of Tygerberg Hospital. Pre-eclamptic patients with acute severe hypertension, who required the placement of brachial intra-arterial lines due to failed radial intra-arterial line placement, were included in the study. Both automated oscillometric and blinded manual aneroid sphygmomanometric blood pressures were recorded during hypertensive peaks and after stabilization of BP using intravenous Labetalol or Nepresol. These two noninvasive methods of blood pressure measurements were compared to brachial intraarterial blood pressure measurements. RESULTS: There was weak correlation between manual and intra-arterial SBP (r = 0.27, p = 0.048) for SBP≥160mmHg. The calculated mean difference between manual SBP compared to the intra-arterial SBP in this group was -23.19mmHg (+/- 19.40). There was moderate correlation between automated and intra-arterial SBP (r = 0.69, p < 0.05). The calculated mean difference between automated SBP compared to the intra-arterial SBP in this group was -16.85mmHg (+/- 11.58). CONCLUSION: This study of pre-eclamptic women demonstrated that both non-invasive methods of BP measurement were inaccurate measures of the true systolic intra-arterial BP and significantly underestimated SBP≥160mmHg when compared to brachial intra-arterial measurements. The SBP was also underestimated, to a lesser degree, for mild moderate hypertension. This study also demonstrated that direct invasive BP monitoring using the brachial artery is a safe method for accurate haemodynamic monitoring. We recommend the use of intra-arterial BP monitoring in pre-eclamptic women with acute severe hypertension. Radial arterial cannulation should be used as the first option and the brachial artery should be used if the first option fails. / AFRIKAANSE OPSOMMING: DOELWIT: Die doel van hierdie studie was om die akuraatheid van nie indringende bragiale bloeddruk metings, wat met outomatiese en manuele aparate geneem is, te vergelyk met bragiale intra-arteriele bloeddruk metings gedurend akute erge hipertensie in pasiente met pre-eklampsie, STUDIE ONTWERP: ʼn Prospektiewe beskrywende dwarssnit studie was in die Obstetriese Kritiese Sorg Eenheid (OCCU) van Tygerberg Hospitaal uit gevoer. Preeklamptiese pasiente met akute erge hipertensie, wat bragiale intra-arteriele lyne nodig gehad het, as gevolg van gefaalde radiale intra-arteriele lyn plasing, was in hierdie studie ingesluit. Beide outomatiese ossilometriese en geblinde aneroide sfigmomanometriese bloeddrukke, tydens hipertensiewe pieke en na stabilisering van bloeddrukke met binneaarse Labetalol of Nepresol, was aangeteken, Die twee nie indringende metodes van bloeddruk meting was met bragiale intra-arteriele bloeddruk metings vergelyk. RESULTATE: Daar was ʼn swak korrelasie tussen manuele en intra-arteriele sistoliese bloedrukke SBP (r = 0.27, p = 0.048) vir SBP≥160mmHg. Die berekende gemiddelde verskil tussen manuele SBP en intra-arteriele SBP was -23.19mmHg (+/- 19.40) in hierdie groep. Daar was ʼn matige korrelasie tussen outomatiese en intra-arteriele SBP (r = 0.69, p < 0.05). Die berekende gemiddelde verskil tussen outomaties SBP vergelyk met intra-arteriele SBP was -16.85mmHg (+/- 11.58) in hierdie groep. GEVOLGTREKKING: Hierdie studie van pre-eklamptiese vrouens, het getoon dat beide nie indringende metodes van bloeddruk meting, nie akurate metings van ware sistoliese intraarteriele bloeddruk is nie, en SBP≥160mmHg word aansienlik onderskat wanneer dit met bragiale intra-arteriele metings vergelyk word. Die SBP was ook tot ʼn minder mate onderskat vir matige hipertensie. Die studie het ook getoon dat die direkte bragiale intra-arteriele metode van bloeddruk monitering, ʼn veilige metode van hemodinamiese monitering is. Ons beveel die gebruik van intra-arteriele bloeddruk monitering aan, in preeklamptiese vrouens met akute erge hipertensie. Radiale arteriele kanulasie moet gebruik word as die eerste opsie en die bragiale arterie moet gebruik word as die eerste opsie faal.