Synthesis, detection and quantification of inulooligosaccharides and fructooligosaccharides by extracellular and intracellular inulinase and fructosyltransferase enzymes isolated from coprophilous fungi.

Masters Degree. University of KwaZulu-Natal, Durban.Exploration of fungal biodiversity capable of producing fructosyltransferase and inulinase enzymes in significant amounts is crucial for the production of oligofructans. Indigenous coprophilous fungi are predominantly sustainable bioresources, harbouring novel enzymes with potential industrial and biotechnological applications. Fructosyltransferase (Ftase) and inulinase are gaining considerable attention due to their capability to synthesise biofunctional nutraceuticals with low calories and health benefits when ingested in recommended dosages. Hence, due to several health benefits associated with prebiotics, bioprospecting for coprophilous fungi as unique bioresources of fructosyltransferase and inulinase was imperative. The present study therefore focused on the collection of herbivore dung from various terrestrial habitats in KwaZulu-Natal Province, South Africa whereby sixty-one (61) indigenous coprophilous fungal strains were isolated after repeated purification to monoculture. The axenic fungal strains were identified using morpho-taxonomic keys and molecular identification by 18S rDNA sequencing where Neocosmospora spp, Trichoderma spp., Aspergillus spp and Fusarium spp. were dominant. The fungal strains were subsequently assessed for their ability to produce extracellular and intracellular Ftase and inulinase enzymes. During the preliminary screening, the culture filtrate was examined for transfructosylating and hydrolytic activity using 2,3,5-triphenyl tetrazolium chloride (TTC) as a chromogenic marker and Lugol’s iodine solution, respectively. Zones of hydrolysis on 30 fungal isolates were observed on the TTC assay plates in diameters ranging from 15 mm to 30 mm, representing high extracellular Ftase activity. The formation of clear zones following addition of iodine solution on inulin rich media indicated the presence of inulinolytic activity. Secondary screening involved DNS assays of eight (8) isolates that secreted high concentrations of Ftase while six (6) different fungal strains showed <50 % inulinase: invertase ratio. The final screening step was tertiary screening where products of biocatalysis were qualitatively detected by thin layer chromatography to visualize saccharide spots of fructooligosaccharides and inulooligosaccharides. HPLC analysis of Ftase and inulinase reaction products revealed and further confirmed that coprophilous fungi harbour fructosyltransferase and inulinase enzymes. The crude extracellular fructosyltransferase enzyme was partially purified by 9.3-fold with a yield of 7.3 % and a specific activity of 2465.5 U mg-1 after a three-step procedure involving (NH4)2SO4 fractionation, dialysis and ion exchange chromatography. The apparent molecular weight of this Ftase was estimated by SDS-PAGE to be approximately 70 kDa. Zymogram analysis under non-reducing conditions showed the enzyme migrating as a polydisperse aggregate yielding broad band of approximately 100 kDa. The enzyme further exhibited an enhanced activity at a broad pH range of 4.0 – 8.0 and optimal activity at a temperature range of 40 °C – 80 °C, while the enzyme was stable at pH 8.0 and between 40 °C – 60 °C, respectively. Under these conditions, the enzyme remained stable retaining 95 % residual activity after incubation for 6 h. The presence of metal ions such as Hg2+ and Ag2+ inhibited Ftase activity while, Ca2+, Mg2+ and K+ at 1 mM increased the enzyme activity, with stabilization observed with Na+, Zn2+ and Cu2+. With sucrose as the substrate, the enzyme kinetics fitted the Michaelis-Menten model. The Km, Vmax and kcat values were 2.076 mM, 4.717 μmole min-1, and 4.7 min-1, respectively with a catalytic efficiency of 2.265 μmole min-1. In vitro antioxidant potential of FOS by 1,1 - diphenyl-2-picryl hydroxyl (DPPH) assay, ferric reducing antioxidant power (FRAP) assay and nitric oxide (NO) radical inhibition yielded IC50 of 6.71 μg/ml, 1.76 μg/ml and IC25 of 0.27 μg/ml, respectively. Free radical scavenging and inhibition activities showed a concentration-dependent antioxidant activity with no significant differences with oligosaccharide standards (p < 0.01). However, vitamin C was significant in FRAP and NO assays. These results clearly demonstrated that an indigenous coprophilous fungus is a potential new reservoir of salient biotechnological enzymes that can be exploited for the production of prebiotics for subsequent biotechnological applications

Impact of Education on Patients’ Perception of Quality Treatment by Male Nurses

ABSTRACT Objective: To improve patients’ perception of quality treatment by male nurses through patient education Design: Quasi-Experimental, Descriptive, Pretest- Posttest Setting: Hays Medical Center, Salina Regional Health Center, and the University of Kansas hospital Participants: Practicing male nurses, male nursing students, and patients Methods: Interviews and questionnaires will be used to cross-examine patients, practicing male nurses, and male nursing students. Education will be provided in order to change patient perception of quality treatment by male nurses Results/Conclusions: Pending results and data collectio

Bone marrow examination findings at aga Khan University Hospital, Nairobi

Objective: To establish the bone marrow examination findings and determine the indication for bone marrow examination. Design: A retrospective audit. Setting: Aga Khan University Hospital, Nairobi. Subjects: All bone marrow aspirates done at Aga Khan University for the period comprising February 2003 to February 2006 were retrieved and analysed. Results: A total of 356 patient’s case histories including bone marrow examination results were analysed. Ages of the patients ranged from 18 months to 91 years. Males were 180 (50.6%) and females were 176 (49.4%). Nutritional anaemia as a group was the most common haematological disorder found on bone marrow examination in our patients with megaloblastic anaemia predominating. Acute myeloid leukaemia was the most common malignant haematological disorder. The most common indication for bone marrow examination was anaemia followed by diagnostic work up of fever of unknown origin. Conclusion: Nutritional anaemia predominated as the commonest benign haematological finding on bone marrow examination while acute myeloid leukaemia was the most frequent haematological malignancy. Most bone marrow examinations were performed on patients with anaemia

Migingo Island Border Dispute in East Africa: A Comparative Analysis of Kenyan and Ugandan Newspaper Coverage

Drawing on agenda-setting, priming and framing theories, this study compares the coverage of Migingo Island ownership dispute in Kenya's Nation and The Standard and Uganda's Monitor and New Vision newspapers. Contents published in 92 issues, between March 1 and December 31, 2009, were studied. The analysis established that both Kenyan and Ugandan newspapers published the conflict mainly in their interior pages with both sides identifying an inexistent borderline as a cause. However, the Kenyan press published the issue more frequently and front-paged a few of its stories while blaming the Ugandan security forces as the main cause of the conflict. Skewed coverage pattern, arising from reporters' nationalistic behavior and desire to serve a domestic audience, was noticed. This has implications for conflict-sensitive reporting of interstate border conflicts in Africa

Urinary tract infections at Aga Khan University hospital Nairobi - A one year experience

Background: In developing countries, most of these patients with urinary tract infections (UTI) are normally treated empirically and urine culture is usual ordered for as a last resort in patients refractory to antibiotic treatment.Objective: To explore the possibility of designing empiric antibiotic therapy for symptomatic UTI in patients at Aga Khan University Hospital by looking at the trends of UTI, common pathogens isolated and their antibiotic susceptibility pattern.Design: A retrospective clinical-laboratory study.Setting: Aga Kahn University Hospital- Nairobi.Subjects: All positive urine cultures between January and December 2008 were included in the study.Results: A total of 409 urine specimens were retrieved and analysed and 100 cases had negative cultures. Three hundred and nine cases had positive cultures. Sixty eight point seven percent were females while 31.3% were males with a mean age of 31 years. One hundred and thirty five out of 409 patients (33%) had classical signs and symptoms. In 143 (35%) cases E. coli was isolated. The other cultures were organisms other than E.coli. There was a higher resistance to clotrimazole in E. coli (71%) as compared to non- E. coli organisms (23%). There was a higher resistance rate to Nalidixic acid in non-E. Coli organisms (35%) and higher resistance rate to Augmentin in E. Coli 43 versus 18% (c/f non-E.Coli). Forty patients in the study had predisposing factors for UTI.Conclusions: It is sometimes warranted to start the patient on empiric antibiotic treatment before culture results are available. Nitrofurantoin, Cefuroxime, Ciprofloxacin have good sensitively rates and are therefore drugs of first choice for the treatment of uncomplicated urinary tract infection provided that the contraindications and specific precautions are noted

Bone marrow examination findings at Aga Khan University Hospital, Nairobi

Objective: To establish the bone marrow examination findings and determine the indication for bone marrow examination.Design: A retrospective audit.Setting: Aga Khan University Hospital, Nairobi.Subjects: All bone marrow aspirates done at Aga Khan University for the period comprising February 2003 to February 2006 were retrieved and analysed.Results: A total of 356 patient’s case histories including bone marrow examination results were analysed. Ages of the patients ranged from 18 months to 91 years. Males were 180 (50.6%) and females were 176 (49.4%). Nutritional anaemia as a group was the most common haematological disorder found on bone marrow examination in our patients with megaloblastic anaemia predominating. Acute myeloid leukaemia was the most common malignant haematological disorder. The most common indication for bone marrow examination was anaemia followed by diagnostic work up of fever of unknown origin.Conclusion: Nutritional anaemia predominated as the commonest benign haematological finding on bone marrow examination while acute myeloid leukaemia was the most frequent haematological malignancy. Most bone marrow examinations were performed on patients with anaemia

Migingo Island Border Dispute in East Africa: A Comparative Analysis of Kenyan and Ugandan Newspaper Coverage

Drawing on agenda-setting, priming and framing theories, this study compares the coverage of Migingo Island ownership dispute in Kenya's Nation and The Standard and Uganda's Monitor and New Vision newspapers. Contents published in 92 issues, between March 1 and December 31, 2009, were studied. The analysis established that both Kenyan and Ugandan newspapers published the conflict mainly in their interior pages with both sides identifying an inexistent borderline as a cause. However, the Kenyan press published the issue more frequently and front-paged a few of its stories while blaming the Ugandan security forces as the main cause of the conflict. Skewed coverage pattern, arising from reporters' nationalistic behavior and desire to serve a domestic audience, was noticed. This has implications for conflict-sensitive reporting of interstate border conflicts in Africa

A Five Year Review OF API20E Bacteria Identification System’s Performance at a Teaching Hospital

Objectives: To assess the performance of the API20E bacteria  identification system at a teaching hospital in Kenya.Design: Retrospective study.Setting: The microbiology laboratoryoratory of the Aga Khan University teaching Hospital.Subjects: One thousand six hundred and fifty eight API20E records.Main outcome measures: The accuracy in identifying the bacteria species.Results: One thousand four hundred and forty two (87.6%) isolates had the exact identity, 199 (12%) nearest identity, and seven (0.4%) no identity. The performance varied among the species; Acinetobacter baumanii had 140 (99.3%) isolates with the exact identity and only one (0.7%) with the nearest identity compared with Aeromonas hydrophila which had five (17.2%) with exact and 24 (82.8%) with nearest.Conclusions: The API20E system is a robust bacteria identification method which can serve small and medium clinical microbiology laboratoryoratories that may not afford automated systems. Adhering to the manufacturer’s instructions and good laboratoryoratory practice can improve the  performance of this method

A realistic explanation of long run development interventions contexts, adaptations and outcomes of dairy improvement in Kenya.

Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2014.Despite continued pursuit of development interventions to improve people’s livelihoods and reduce poverty, intended and actual outcomes of developments interventions may differ. Some scholars attribute this variation to flawed conceptualisation of development interventions while others view this divergence as evidence that implementation processes are complex and actual outcomes result from adaptations of the interventions by actors. To move beyond the discursive approaches to analysis of development interventions, this thesis addresses the question how do actors adapt them, why and with what outcomes in the long run? Empirically, it looks at how project officers and farmers adapted the National Dairy Development Project (NDDP), a dairy intervention implemented in Kenya between 1980 and 1995, and its long run outcomes. The intervention promoted zero grazing, intensive management of dairy cattle whose implementation by farmers was expected to increase land productivity as a means to address land scarcity, increase milk production and reduce poverty through generation of incomes from milk sales. The methodology of this thesis links mechanisms, contexts, and outcomes, three elements of realist explanation, to understand adaptations and outcomes of development interventions. Through thematic synthesis of in-depth interviews and analysis of project documents, this thesis explains adaptations and long run outcomes of the NDDP. Findings reveal that developers and farmers adapted several components of the intervention. With close reference to context, incentives and continuity pressures, this thesis utilises intervention effectiveness and matching mechanisms to explain how project officers adapted the NDDP. Further, through fit and resistance mechanisms, this thesis explains how farmers adapted zero grazing in the context of inadequate fodder, labour shortage and lack of resources to invest in dairy. In the long run, findings show that the intervention diminished as evident in coexistence of indigenous and modern dairy technologies and non-implementation of any dairy technologies by farmers. Despite adaptations of zero grazing by project officers and farmers, intensification of dairy cattle management has diminished in the context of resource constraints, neoliberal policies and labour shortage. Consequently, the objective to increase land productivity through intensive dairy cattle management, the rationale for initiation of the intervention, remains unresolved

Resilience in a Kenyan informal settlement during the COVID-19 pandemic.

Informal settlements have been identified as locations both where the spread of COVID-19 has generally been slower than within the Global North and measures to restrain the pandemic have further intensified local peoples’ marginality as income decreases without welfare or financial safety nets. In this paper, qualitative fieldwork is detailed which commenced in Korogocho, an informal settlement in Nairobi, Kenya, immediately prior to national COVID-19 restrictions. This March 2020, pre-Covid phase of the fieldwork, focused on a community based project and the basis for resilience in transforming local lives. During the next 12 months of the pandemic fieldwork continued, exploring experiences and reactions to restraining policies. These findings reinforce concerns about the impact of COVID-19 related restrictions on marginalised peoples’ income, food security, health, safety and gender based violence. How the local people reacted to these effects highlights their creative resilience and adaptability. The paper concludes by examining the impact of, and responses to, the controlling measures on the social relationships and cohesion that underpins the community resilience