Thesis (MMed)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Background Occult blood loss from the gastrointestinal tract (GIT), causing iron deficiency often with anaemia, can be diagnostically and therapeutically challenging. This is because the endoscopic and radiologic tests may be negative due to the slow, chronic and intermittent nature of the gastrointestinal bleeding, making timing key in detection and localisation of the bleed. These limitations can be approached using two different radioactive isotopes. Firstly, we tested the sensitivity of extending Cr-51 RBC for 21 days relative to 5 days to detect GIT bleeding and its use to optimise timing of a Tc-99m RBC study for GIT blood loss localisation. Finally, we tested if the information provided by the Tc-99m RBC study aided gastroenterologic intervention for anatomical localisation of a lesion. Method In this retrospective review, after obtaining institutional and ethics committee approval, records of patients referred for evaluation of possible GIT blood loss were reviewed. In each; daily appearance of radiochromium in stool was measured in the whole body counter. In those cases exceeding 50 ml/day, a technetium-99m (Tc-99m) localization study was performed. These studies were correlated with clinical findings. Results A total of 59 Cr-51 RBC studies were carried out in 36 females and 21 males (n = 57). In 32 (54%) the radiochromium results were positive with 75% of the bleeding incidences occurring after 5 days of stool collection. Of 17 cases in whom Tc-99m RBC imaging studies were performed, 14 (82%) were positive with specific anatomical sites successfully defined in twelve. In all patients with blood loss of >100 ml/24h, Tc-99m RBC were positive and localised. Ten of the 17 Tc-99m RBC studies were further investigated and half diagnosed with small-bowel angiodysplasia. Conclusion This sequential twin isotope method is practical in revealing otherwise silent intestinal haemorrhage. Although it has good patient acceptability and clinical as well as diagnostic utility in management, further studies are required to clearly establish a cut-off level of blood loss for performing imaging studies and the impact of the findings on the overall patient management. / AFRIKAANSE OPSOMMING: Agtergrond Die evaluasie van okkulte bloedverlies uit die gastro-intestinale kanaal (GIT), met gevolglike ystertekort anemie, kan diagnosties en terapeuties uitdagend wees. Dit is omdat endoskopiese en radiologiese ondersoeke negatief mag wees as gevolg van die stadige, chroniese en intermitterende aard van die gastro-intestinale bloeding, wat die presiese tydstip van opsporing en lokalisering van die bloeding krities belangrik maak. Hierdie beperkings kan aangespreek word deur twee verskillende radioaktiewe isotope te gebruik. Eerstens is die sensitiwiteit van die verlenging van die Cr-51 RBS studie tot 21 dae in plaas van 5 dae om die GIT bloeding op te spoor, getoets, asook die gebruik daarvan om die optimale tyd vir ‘n Tc-99m RBS studie om die GIT bloedverlies te lokaliseer, vas te stel. Laastens is getoets of die inligting van die Tc-99m RBS studie wel bygedra het tot die gastroenterologiese ingreep om die letsel anatomies te lokaliseer. Metode Na institusionele en etiese komitee toestemming is inligting van pasiënte wat vir die evaluering van ‘n moontlike GI bloedverlies verwys is, in hierdie retrospektiewe oorsig nagegaan. Die daaglikse voorkoms van radioaktiewe chroom in stoelgangmonsters is in ‘n heelliggaamteller gemeet. In gevalle waar dit 50 ml/dag oorskry het, is ‘n tegnesium 99m (Tc 99m) studie gedoen. Hierdie studies is met die kliniese bevindinge gekorreleer. Resultate ‘n Totaal van 59 Cr-51 RBS studies is in 36 vroue en 21 mans (n = 57) gedoen. Die gemerkte chroomstudies was positief in 32 (54%), met 75% van die bloedings wat meer as 5 dae na versameling van die stoelgang plaasgevind het. In veertien (82%) van die 17 gevalle waar Tc-99m RBS studies gedoen is, was die studies positief. Spesifieke anatomiese gebiede van bloeding kon in 12 hiervan suksesvol bevestig word. Tc-99m RBS studies was positief in al die pasiënte met ‘n bloedverlies van >100 ml/24h, en kon gelokaliseer word. Tien van die 17 Tc-99m RBS studies is verder ondersoek en die helfte daarvan gediagnoseer met dunderm angiodisplasie. Gevolgtrekking Die opeenvolgende twee isotoopmetode om andersins asimptomatiese dermbloeding op te spoor, is prakties uitvoerbaar. Alhoewel die studies goed deur pasiënte aanvaar is, en ook van kliniese en diagnostiese waarde in die hantering van die pasiënte is, is verdere studies nodig om die afsnypunt vir die hoeveelheid bloedverlies om beeldingstudies uit te voer, sonder twyfel vas te stel, asook om die impak van die bevindings op ‘n groter pasiëntpopulasie vas te stel.

In dieser Studie wurde eine Charakterisierung von Eisenpräparaten entwickelt, die als parenterale und orale Präparate zur Behandlung schwerer Anämie auf Basis von Fe-Kohlenhydrat-Nanopartikeln und Bioverfügbarkeitsstudien in biologischen Proben verwendet werden. Eine vollständige Charakterisierung eines kommerziellen Präparats für die parenterale Behandlung von Anämie (Venofer®) wurde durchgeführt. Solubilisationsstudien des Arzneimittels wurden in Serum und Blut durchgeführt, um die Bioverfügbarkeit von Eisen zu untersuchen. Es wurden Speziationsexperimente von Serumproteinen durchgeführt (UV-vis, ICP-MS), die detailliertere Informationen über das Arzneimittel und sein Verhalten im Körper liefern. Tartrat-modifizierte Eisenoxid-Nanopartikel (FeNPs) und isotopisch angereicherte 57Fe wurden synthetisiert. Nanopartikel wurden weiter in Bezug auf Partikelgröße und -form charakterisiert. Da diese Nanopartikel möglicherweise für die orale Supplementierung bei der Die Labilität dieser FeNPs wurde in saurem Medium bewertet, und das gesamte Fe wurde in nanopartikulärer und löslicher Fraktion quantifiziert. Zur Bestimmung des Nanostrukturverhaltens in zellulären Medien wurde eine Speziationsstrategie für Nanopartikel und ionische Eisenspezies basierend auf einer reversed phase high performance liquid chromatography, gekoppelt an ICP-MS, durchgeführt und erstmals auf Caco-2-Zellen angewendet Proben. Darüber hinaus wurde ein Quantifizierungsansatz durch Anwendung einer Isotopenverdünnungsanalyse mit 57Fe. Die Zytotoxizität wurde untersucht: kolorimetrische Assays wurden in verschiedenen Zelllinien durchgeführt, und die oxidative Schädigung wurde durch Messung der ROS-Produktion bestimmt. Schließlich wurden In-vivo-Modelle zur Bestimmung der Eisenresorption im Dünndarm nach Perfusionsexperimenten an Ratten verwendet. Das Schicksal der FeNPs wurde auch untersucht, indem andere Gewebe wie Leber, Niere, Milz oder Blut nach der intestinalen Aufnahme analysiert wurden. / In this study, a characterization of iron supplements used as parenteral and oral preparations for the treatment of severe anaemia based on Fe-carbohydrate nanoparticles and bioavailability studies in biological samples have been developed. A complete characterization of a commercial preparation for the parenteral treatment of anaemia (Venofer®), has been carried out. Solubilisation studies of the drug have been conducted in serum and blood to evaluate iron bioavailability. Moreover, speciation experiments of serum proteins were conducted using UV-vis and ICP-MS detection, providing more in-depth information about the drug and its behaviour in the body. Tartrate-modified coated-iron oxide nanoparticles (FeNPs) and isotopically enriched 57FeNPs have been synthesized. Nanoparticles were further characterized in terms of particle size and shape. As these nanoparticles are potentially suitable for the oral supplementation in anaemia’s treatment, the lability of these FeNPs in acidic medium was evaluated, and total Fe was quantified in nanoparticulate and soluble fraction. For the determination of nanostructure behaviour in cellular media, a speciation strategy for nanoparticles and ionic species of iron based on a reversed-phase high-performance liquid chromatography coupled to ICP-MS was carried out and applied for the first time to Caco-2 cell samples. Moreover, a quantification approach was also tackled by applying an online post-column isotope dilution analysis with 57Fe as isotopically enriched standard. Cytotoxicity from these tartrate-modified iron oxide nanoparticles was evaluated: colorimetric assays have been conducted in different cell lines and oxidative damage was assessed by measuring ROS production. Finally, in vivo models were used to determine iron absorption in the small intestine after perfusion experiments in rats. The fate of the FeNPs was also studied by analysing other tissues as liver, kidney, spleen, or blood after the intestinal uptake.

This parallel randomized control trial study assessed the effect of food-based strategies in improving the haemoglobin level; decreasing anaemia and thus the iron status of pregnant women. The study randomized 195 anaemic women enrolled from four randomly selected health centres in Dire Dawa while attending ANC into two intervention groups and control; and followed for 12weeks. The study intervened diet-based supplementation of 90mg/day vitamin C divided and consumed in three doses; combined with nutrition education intervention in intervention group 1; while only nutrition education intervention in group 2. Dietary diversity was assessed using past 24-hours and 7-days dietary recall approaches and haemoglobin levels were determined at baseline and end-line and compared to analyse treatment effects. By end of the study, intervention group 1 and 2 had significantly increased mean haemoglobin by 0.77 ± 0.11gm/dl and 0.398 ± 0.073gm/dl respectively; however, the control had significant decrease by -0.193 ± 0.05gm/dl. Anaemia prevalence also significantly decreased by 29% and 19.7% in intervention group 1 and 2 respectively. However, all women in the control were anaemic. Intervention group 1 and 2 also had significantly higher dietary diversity, consumption of vitamin C-rich fruits and vegetables, nutritional knowledge and modification practices. The researcher thus concludes that diet-based vitamin C supplementation integrated with nutrition education has a significant effect in improving haemoglobin, decreasing anaemia and thus improving the iron status of pregnant women in Dire Dawa. Based on the findings, the researcher developed a framework for an integrated food-based strategy for improving the iron status of pregnant women in Ethiopia. / Health Studies / D. Litt. et Phil. (Health Studies)