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Introduction: Bronchiectasis is a common chronic debilitating respiratory condition. Patients suffer daily cough, excess sputum production and recurrent chest infections because of inflamed and permanently damaged airways. The pathogenesis of bronchiectasis is poorly understood. Pulmonary pathology shows excess neutrophilic airways inflammation, but despite this over two thirds of patients are chronically infected with potential pathogenic microorganisms. The acute inflammatory response is a protective mechanism that is evolved to eliminate invading organisms and should ideally be self-limiting and lead to complete resolution. The driver for persistent neutrophilic airway inflammation in bronchiectasis is unknown, but infection is considered to play a major role. AIMS The main aims of this thesis were to: (i) Characterize neutrophils in the serum and airways in bronchiectasis in the stable state and during exacerbations; (ii) Cohort study to establish if LXA4 deficiency correlates with disease severity (iii) Characterize lipids in bronchiectasis airways and peripheral blood to establish the correlation of LXA4 to disease severity; (iv) To investigate a potential mechanism for low levels of LXA4 in bronchiectasis, lipoxin biosynthetic genes expression will be measured; (v) Assess the anti-inflammatory and pro resolution effect of LXA4 on neutrophils and monocyte derived macrophages from healthy volunteers; (vi) Assess the anti-inflammatory and pro resolution effect of LXA4 on neutrophils during exacerbations in bronchiectasis and community acquired pneumonia. Methods (I) To establish the serum neutrophil subtype in stable state and following antibiotic treatment in patients with bronchiectasis, the following studies were done. Inclusion criteria: Patients aged 18-80 were recruited. All had an established radiological diagnosis of bronchiectasis (CT of the chest). Patients had clinically significant bronchiectasis, aetiology being either idiopathic or post infection. Exclusion Criteria: current smokers or ex-smokers of less than 1 year; >20 pack year history; cystic fibrosis; active allergic bronchopulmonary aspergillosis; active tuberculosis; poorly controlled asthma; severe COPD requiring nebulised bronchodilators or long term oxygen therapy; patients on aspirin or leukotriene inhibitors, pregnancy or breast feeding, active malignancy. A. 6 patients with mild bronchiectasis, 6 patients with severe bronchiectasis and 6 healthy volunteers were recruited. Serum and airways neutrophils were subsequently isolated. Neutrophil apoptosis, CD11b and CD62L expression, myeloperoxidase release, superoxide generation, phagocytosis and killing of GFP labeled bacteria were assessed. B. To compare serum with airways neutrophils function, bacterial phagocytosis and killing of GFP labeled bacteria was done, with both serum and airways neutrophils. Samples were obtained from the above group of patients. C. To establish neutrophil function following antibiotic treatment, 6 bronchiectasis patients at the beginning (day1) and the end (day14) of intravenous antibiotic therapy for an exacerbation were studied. As a control group, 6 community acquired pneumonia patients at the beginning (day1) and the end (day 5) of intravenous antibiotic therapy for infection were studied. Induced sputum and peripheral blood was taken at day1 and 5, where able. Phagocytosis and killing of GFP labeled bacteria was assessed and the two groups compared. (II) To address if lipoxin A4 deficiency correlates with disease severity, a cohort study was done in bronchiectasis patients. 169 patients were recruited and followed up for 1 year. Assessments done were Bronchiectasis severity index, systemic inflammatory markers (white cell count, ESR and c-reactive protein), Forced Expired Volume in 1sec, Forced Vital Capacity and its ratio, antibiotic courses in 1 year, hospital admissions in 1 year, sputum microbiology, quality of life assessments by Leicester Cough Questionnaire and St. Georges Respiratory Questionnaire, interleukin 8, myeloperoxidase, neutrophil elastase and leukotriene B4 (from sputum). (III) To assess effect of lipoxin on disease severity, 6 healthy volunteers, 10 patients with mild disease, 15 with moderate and 9 with severe disease were recruited. Disease severity was calculated as per the bronchiectasis severity index. All participants had 60mls of blood taken and underwent a bronchoscopy. Two segments of the lungs were washed out from bronchiectasis patients, an area affected by bronchiectasis and an area unaffected by bronchiectasis. This led to patients acting as their own internal control. Serum and airways neutrophils (from both segments) were subsequently isolated. Assessments done were systemic inflammatory markers (white cell count, ESR and c-reactive protein), serum lipoxin A4 and the cathelicidin LL-37, Forced Expired Volume in 1sec, Forced Vital Capacity and its ratio, transfer factor for carbon monoxide, antibiotic courses in 1 year, hospital admissions in 1 year and sputum microbiology. Phagocytosis and bacterial killing were assessed by both serum and airways neutrophils. From bronchoalveolar lavage fluid (BALF), I measured myeloperoxidase and neutrophil elastase. For both serum and BALF, lipidomics were obtained. (IV) To address the impact of anatomic compartment, gene expression was measured in from endobronchial brushings from the same cohort of bronchiectasis patients and controls as above, where samples were available. qPCR was performed for the following eicosanoid biosynthetic genes- 5 Lipoxygenase (LOX), 15 LO-A, 15LO-B and leukotriene (LT) A4 hydrolase. (V) To assess the anti inflammatory and pro resolution effect of LXA4 on neutrophils and monocyte derived macrophages from healthy volunteers, freshly isolated PMN will be treated with LXA4 or vehicle control. Spontaneous apoptosis was measured. fMLF and cytochalasin B was added and the inflammatory response assessed measuring myeloperoxidase (MPO), free neutrophil elastase (NE), CD11b, CD18 and CD62L. Human monocytes and PMNs were isolated from bronchiectasis patients. Following differentiation, LXA4 treated or control adherent, washed MDMs will be incubated with apoptotic stained PMNs. Efferocytosis was analyzed by flow cytometry. (VI) To establish the effect of Lipoxin A4 on neutrophil function following antibiotic treatment, the same study group used to evaluate aim 1 was taken. As a control group, 6 community acquired pneumonia patients at the beginning (day1) and the end (day 5) of oral or intravenous antibiotic therapy for infection were studied. Induced sputum and peripheral blood was taken at day1 and 5, where able. Phagocytosis and killing of GFP labeled bacteria and the effect of Lipoxin A4 was assessed and the two groups compared. Serum and sputum lipidomics were obtained in bronchiectasis exacerbations on day 1 and day 14. Serum lipidomics was obtained in pneumonia on day 1 and day 5. RESULTS (I) Neutrophil sub type study (Studied on healthy volunteers/ mild/ severe bronchiectasis) Peripheral blood neutrophils from bronchiectasis patients showed that there was significantly more viable neutrophils in mild and severe bronchiectasis compared to healthy volunteers, p=0.002 and p=0.005 respectively. In addition, there was significantly less apoptotic neutrophils in mild and severe bronchiectasis compared to healthy volunteers, p=0.0003 and p < 0.0001 respectively. There was a significantly higher level of CD11b in the mild (p=0.01) and severe bronchiectasis (p=0.01) compared to healthy volunteers. There was more CD62L shedding (p=0.02) and myeloperoxidase release (p=0.04) in bronchiectasis compared to healthy volunteers. There was lesser phagocytosis in mild (p=0.04) and severe (p=0.03) bronchiectasis compared to healthy volunteers. This led to lesser bacterial killing in mild (p=0.04) and severe (p=0.0004) bronchiectasis compared to healthy volunteers.

The purpose of this research was to determine the relationship of the slime layer antigen(s) to the "101" or LPS antigens and to attempt to evaluate the role of antibodies against the latter in protection against experimental infections in mice with the homologous strain of Pseudomonas aeruginosa. Results from agglutination tests, chromatographic separations, passive protection tests, and characterizations of the antigens by gel double diffusion do not support the concept that LPS is a necessary portion of the immunogenic material. The immunogenicity of LPS can be attributed to co-purification of residual amounts of slime layer antigens on the washed cells from which LPS was extracted.

Thesis (MSc. Agriculture (Agronomy)) --University of Limpopo, 2015 / Agronomic field trial was planted at two sites, Ukulima and Syferkuil, in Limpopo Province, to determine the response of cowpea to variable rates and methods of zinc application under supplementary irrigation and rainfed conditions, respectively. The experiment was laid out in a split plot arrangement and fitted into a randomized complete block design (RCBD). Treatments consisted of two factors namely zinc rates (0, 5, 10, 15, 20 and 25 kg ha-1) and methods of application (soil and foliar) with white cowpea variety (IT00K-1217) seeds sown. The zinc fertilizer was applied as ZnSO4 (38.5% Zn) while basal phosphorus (30 kg P ha-1) was applied in the form of single super phosphate (10.5% P) so as to eliminate P constraints. Soil application of the zinc fertilizer was band placed near the row of cowpea plant while foliar application was done 3-5 weeks after plant emergence. Growth parameters measured included plant height, number of primary branches, canopy cover, number of trifoliate leaves, days to 50% flowering and pod formation, and fresh biomass as well as yield component attributes. Twenty young cowpea leaves and immature green pods were each harvested at approximately 75% physiological maturity. Crude protein, total nitrogen, potassium, phosphorus, zinc, and iron content and uptake were assessed in the different plant parts. Growth, yield and nutrients data collected were subjected to analysis of variance. Treatments means were separated using Tukey’s test at probability level of 5%, while the response variables were modelled using quadratic polynomial equation. Results obtained revealed that total above ground biomass yield of 6219 kg ha-1 and fresh pod weight of 142.3 g were obtained when zinc was soil applied under rainfed condition compared to 6019 kg ha-1 and 138.0 g for total above ground biomass yield and fresh pod weight, respectively when foliar applied. Soil application gave a total above ground biomass yield of 6298 kg ha-1 and pod fresh weight of 150.9 g while foliar application gave total above ground biomass yield and pod fresh weight of 4791 kg ha-1 and 124.0 g, respectively at Ukulima. Soil application also gave a significantly higher grain yield (2251 kg ha-1) than the foliar application (1503 kg ha-1) at Ukulima. A higher but inconsequential effect on grain yield was obtained with soil over foliar application at Syferkuil. Application of zinc fertilizer at a rate beyond 5 kg ha-1 resulted in a grain yield reduction of up to 22.2% and 6.6% respectively at vi Ukulima and Syferkuil. Thus, this rate appeared optimum for both grain and fodder production when soil applied at both sites. Based on the quadratic model, total biomass (4897 kg ha-1) and grain (1602 kg ha-1) yields were optimized at an estimated zinc rate of 54.7 and 33.4 kg ha-1, respectively under supplementary irrigation, while total biomass (5913 kg ha-1) and grain (2696 kg ha-1) yields were obtained at an estimated optimum zinc rate of 20.1 and 26.8 kg ha-1, respectively under rainfed condition. A scorching effect of cowpea leaves was observed following foliar application of zinc fertiliser at 25 kg ha-1 that resulted in a decreased growth and yield at Ukulima. Foliar application resulted in improve cowpea leaf zinc concentration (43.9 mg kg-1) compared to soil application (23.2 mg kg-1) at Syferkuil. It also resulted in increased crude protein, total N, P, Fe and Zn uptake compared to soil application at Ukulima. Both soil and foliar zinc application gave 28.5% zinc concentration in the fresh immature pods samples collected. Findings from this study reveal that foliar zinc sulphate application resulted in higher grain and biomass yields as well as zinc concentration in the various cowpea plant parts. Based on the quadratic model used in this study, the optimum zinc rate required to guarantee high cowpea yield and zinc-rich grain and leaf content at both trial sites is highly variable; and thus requires further study for validation. Keywords: Cowpea production, grain yield, dryland farming, leafy vegetables, foliar fertilizer, improved nutrition, zinc deficiency.

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Papua New Guinea has been classified by the World Health Organisation as an area where clinical vitamin A deficiency (VAD) exists. This is at variance with the experience of the local physicians who do not encounter classical VAD in clinical practice. This pilot study was carried out to resolve this contradiction, since many suggestions have been made to fortify foods with Vitamin A. If done in the absence of concrete data to back up this classification, it could take the focus away from the real problem as well as potentially create problems of Vitamin A toxicity. Therefore, answers from this study could have far reaching implications in a country such as PNG, which has high childhood mortality and limited financial and manpower resources. The objective of this study was to determine the vitamin A status and identify risk factors of VAD in children aged 6 months to 6 years in a rural area of Papua New Guinea. The survey was carried out in the Ramu region of Madang province. Households and children were randomly selected and standard questionnaires were used to collect information about diet, previous illnesses and night blindness. The weight and height of all children was recorded and an ocular and physical examination carried out by trained personnel. In addition, haemoglobin estimation and examination of blood films for malaria parasites was carried out in all the children. In a randomly selected number of children, estimations of serum retinol and other micronutrient levels were carried out. Results: A total of 609 children were enrolled in the study. Biochemical parameters were studied in 106 of them .The mean age of the children was 35 months. Possible night blindness was reported in 4 children. No xerophthalmia was seen. The prevalence of serum retinol levels ( 0.7 (mol/L (WHO suggested cut off values for subclinical VAD) was 10.3%. Anthropometric indicators indicated a high proportion of the children had stunting and wasting or both. Analysis of dietary patterns, maternal literacy, food availability and other surrogate indicators indicated that the population is at mild-moderate risk of developing VAD. In conclusion, no evidence of clinical vitamin A deficiency was found. Subclinical vitamin A deficiency seemed to occur in this population at a level of mild-moderate public health importance. Further studies need to be carried out to assess the situation in different areas in PNG before policy decisions can be made with regards to mass vitamin A supplementation.

This thesis describes an investigation of the effects of vitamin A deficiency on gut function, The central hypothesis to be tested was that acute vitamin A deficiency affects glucose uptake from the small intestine- The hypothesis was tested using a system involving perfusion of isolated segments of the small intestine in the anaesthetized rat. The system was used to study effects on glucose uptake under steady-state conditions. In the initial part of the study, experiments were diverted towards setting up the system for measuring steady-state uptake, and determining the relative contributions of active uptake and diffusion. Phenol red was found to be a reliable non-absorbable marker for determining net water movement. Phlorizin, generally at 1 mmol/L, was used as a competitive (reversible) inhibitor of active uptake. It is difficult however to confirm complete inhibition of active uptake by phlorizin because of the limited solubility of the inhibitor. The kinetics of glucose uptake f ram intra-luminal maltose were found to be, in general, not significantly different from those applying to the uptake of glucose from an equivalent glucose solution. Maltase activity in the perfused gut segment was found to be sufficient to hydrolyse most of the maltose (80 per cent or more) in the solution being perfused, a much greater proportion than was absorbed. Glucose absorptive capacity, measured on an intestinal dry weight basis, was greatest in the duodenum and progressively less in the jejunum and ileum. The rate of water uptake f ran the gut was increased by the presence of glucose in the lumen, and was linked to glucose uptake as shown by the inhibition of water uptake by phlorizin. Uptake of glucose by solvent drag was demonstrated by showing an increased rate of glucose uptake when the rate of water uptake was increased by perfusing a solution of reduced osmotic pressure. In the experiment a low intra-luminal glucose concentration was used to preclude net uptake by diffusion and active uptake was blocked with phlorizin. This process was further investigated using streptozotocin-diabetic rats in which the diabetes establishes a hyperosomotic blood with hyperglycaemia. Uptake by solvent drag was more obvious in diabetic animals. A back-diffusion (exsorption) of glucose from the tissues to the lumen was also shown; the rate being proportional to plasma glucose concentration. Vitamin A deficiency was established in weanling rats after 6-7 weeks feeding on a diet based on wheat starch, coconut oil, and casein washed with hot ethanol, together with vitamins and minerals. The vitamin A deficiency led to classic eye signs and was reversed by the addition to the diet of retinoic acid (5 g/g diet). Vitamin A deficiency decreased intestinal mucus production (dry weight) but had no detectable effect on the histology of the villous epithelium as shown under the light microscope. Using perfusion experiments it was shown that vitamin A deficiency had no significant effect on the rate of active uptake of glucose, but that deficiency increased the rate of passive uptake.

Bibliography: leaves 218-233. / xiii, 233 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This thesis identifies a strong relationship between Zn nutrition of medics and severity of disease by R. solani (AG8) which may be a major factor contributing to the current decline in medic pastures observed in Australia. The effect of seed nutritional value had a profound impact on the growth of medics and may also be a key factor in poor medic growth. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 1998

Study Objective. To assess the dietary intakes of children with Attention Deficit Hyperactivity Disorder (ADHD); including macro- and micronutrient intake, the percentage of low nutrient density foods in the diet, as well as specific biochemical and anthropometric parameters. Design. Observational study. Setting. Provincial ADHD Program of BC, Children’s’ Hospital, Vancouver, BC. Sample. 44 children aged 6 - 12 years with ADHD, treatment-naïve or stable on medication for 6 months or longer. Main Results. ADHD Children were taller and heavier than population norms, and significantly taller than previously reported in ADHD. Of children aged 6-8 years, 28% were below the Estimate Average Requirement (EAR) for zinc and 61% of children aged 9-12 years were below the EAR for zinc. In addition, 28% of children aged 6-8 years and 39% of children aged 9-12 years were below the EAR for copper. Only 2% of children in the sample were below lab normal cutoffs for ferritin; however, 73% had serum zinc values below lab normal cut-offs and 23% had serum copper below lab normal cutoffs. Overall prevalence of serum zinc below the 2.5 percentile was 23% compared to 2% from National Health and Nutrition Examination Survey II (NHANES) data. Additionally, mean serum copper was significantly lower than NHANES II data. Mean energy intake was comparable to mean Estimated Energy Requirement (EER) based on the age, gender, height, weight and physical activity of subjects. In addition, mean dietary intake of Low Nutrient Density foods was not significantly different from NHANES II data and there was no significant difference in energy intake or the proportion of daily energy from protein, fat, and carbohydrate than what is observed from CCHS data. Medication treatment for ADHD was not associated with altered dietary intake or nutrient status. Conclusion. Results are suggestive of low zinc status in ADHD.

Maternal and child undernutrition are a significant problem in the developing world, with serious consequences for human health and socio-economic development. In Cambodia, 55% of children, 43% of women of reproductive age, and 50% of pregnant women are anemic. Current prevention and control practices rely on supplementation with iron pills or large-scale food fortification, neither of which are affordable or feasible in rural Cambodia. In the study areas, 97% of women did not meet their daily iron requirements. The current research focuses on the design and evaluation of an innovative iron supplementation technique. A culturally acceptable, inexpensive and lightweight iron ingot was designed to resemble a fish species considered lucky in Khmer culture. The ingot, referred to as ‘try sabay’ or ‘happy fish’, was designed to supply iron at a slow, steady rate. Iron leaching was observed in water and soup samples prepared with the iron fish when used concurrently with an acidifier. More than 75% of daily iron requirements can be met with regular use. Its use in the common pot of soup or boiled water provides supplementation to the entire family. The effectiveness of the iron fish was investigated in a randomized community trial involving 310 women in rural Cambodia. Blood samples were taken at baseline and every three months thereafter, over a 12-month trial period. Significant increases in hemoglobin concentrations were observed in women allocated an iron fish when compared to controls throughout the study, with an endline difference of 11.6 g/L. Significant improvements in serum ferritin concentration were observed at 9 months (6.9 ng/mL) and endline (30.8 ng/mL) in women who used an iron fish regularly when compared to the control group. Overall, use of the iron fish led to a two-fold reduction in the prevalence of anemia. The supplement was used daily by 94% of the households at the end of the trial. The study highlights the acceptability and effectiveness of a fish-shaped iron ingot as a means of improving dietary iron content. It offers a promising, simple solution to iron deficiency anemia if the project can be scaled-up for use throughout the country. / Canadian Institutes of Health Research, University of Guelph, International Development Research Centre (Canada)