Photophysicochemical properties and in vitro photodynamic therapy activities of zinc phthalocyanine conjugates with biomolecules and single-walled carbon nanotubes
- Authors: Ogbodu, Racheal O
- Date: 2015
- Subjects: Photochemotherapy , Phthalocyanines , Biomolecules
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4544 , http://hdl.handle.net/10962/d1017924
- Description: The synthesis, photophysicochemcial properties, in vitro dark toxicity and photodynamic therapy (PDT) activities of different derivatives of zinc phthalocyanine (ZnPc) conjugates with biomolecules (folic acid, bovine serum albumin (BSA), ascorbic acid, uridine or spermine) and single-walled carbon nanotubes (SWCNTs) are presented in this work. The fluorescence quantum yields (ΦF) (Subscript F) of the ZnPc derivatives or ZnPc-biomolecule conjugates remained relatively the same as compared to the precursor Pcs. Slight increases were observed in the ΦF (Subscript F) values of conjugates containing substituents such as pyrene, folic acid or BSA with intrinsic fluorescence properties. The triplet quantum yield (ΦT ) (Subscript T) values for some ZnPc conjugates increases compared to the precursor ZnPcs due to extended π conjugation (for the conjugate with pyrene) and the presence of phenyl ring that support spin-orbit charge transfer intersystem crossing to triplet state. While some conjugates showed decreases in the ΦT (Subscript T) values compared to precursor ZnPcs due to the presence of substituents that could quench photo-excited state properties. The singlet oxygen quantum yield (ΦΔ ) values follow the trends of the triplet quantum yield values. The conjugates containing BSA also show increases in the ΦΔ values without corresponding increases in ΦT (Subscript T) values due to the ability of BSA to generate free radicals including singlet oxygen. The presence of SWCNTs decreases the photophysicochemcial properties of some ZnPc-SWCNT conjugates compared to the precursor ZnPcs due to photo-induced electron transfer from an excited Pc complex (electron donor) to SWCNTs (electron acceptor). However, increases were observed in some ZnPc-SWCNT conjugates as a result of fast charge recombination process due to highly short-lived radical ion pair produced. These phenomena affected the ΦF (Suscript F) values, ΦT (Suscript T) values, and the ΦΔ values. Increases or decreases in ΦT (Suscript T) values resulted in corresponding increases or decreases in ΦΔ values
- Full Text:
- Date Issued: 2015
The design, synthesis and antiplasmodial activity of a series of halogenated fosmidomycin analogues and hybrid drugs
- Authors: Afolayan, Anthonia Folake
- Date: 2012
- Subjects: Uncatalogued
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64370 , vital:28538
- Description: Malaria continues to be a devastating disease and a major cause of death in sub-Saharan Africa. With resistance against most of the available antimalarial drugs, there is a need for ongoing research and development of antimalarial agents. Fosmidomycin and its acetyl analogue FR900098 have been identified as potent inhibitors of Plasmodium falciparum, the causative agent of the most deadly form of malaria. Clinical trials of these agents have revealed poor absorption due to their high hydrophilicity. In the present studies the effect of halogenation of the acyl chain as well as the biological effect of extending the acyl sidechain was explored. This provided the basis on which fosmidomycin hybrids were designed to investigate the feasibility of hybrid extending into NADPH binding pocket. Synthesis of a series of halogenated FR900098 analogues was carried out in three stages. This included i) The introduction of the phosphonate group by reaction with 1,3dibromopropane in an Arbuzov reaction, ii) The introduction of a hydroxamate group by reaction of the propyl phosphonate by means of a nucleophilic substitution reaction with BocNHOBn and iii) The introduction of a halogenated acyl side chain on a protected fosmidomycin backbone. The synthesis of fosmidomycin-hybrids for which chloroquinefosmidomycin hybrids were used as the prototype, involved convergence of the two separately constructed moieties i.e. fosmidomycin and the quinoline moieties in a covalent linkage. The quinoline moiety was easily synthesized from the reaction of 4,7dichloroquinoline with 1,2-diamino ethane. The aminoquinoline so formed resulted in chloroquine-fosmidomycin hybrids 3.8 and 3.9 when reacted with halogenated FR900098 analogues. Antiplasmodial assays were conducted on the chloroquine-fosmidomycin hybrids and the halogenated fosmidomycin derivatives against the chloroquine resistant Gambian FCR-3 strain of P. falciparum. The most potent iodoacetyl fosmidomycin analogues 2.21 gave an IC50 value of 5.54 µM which is eight times more potent than the known antiplasmodial FR900098 which gave an IC50 value of 41.67 µM. All the halogenated FR900098 analogues showed better antiplasmodial activity than their non-halogenated derivatives. This indicated that the presence of halogens in the FR900098 analogues contributes to their biological Chapter 1 Literature review activity. The acetyl and propyl linked hybrids 3.8 and 3.9 showed potent antiplasmodial activity with IC50 values of 0.18 and 0.82 µM respectively. These were by far the most potent hybrids synthesized and provided leads for a new class of promising antimalarial agents. Preliminary E. coli DXR enzyme inhibition assays were carried out on the halogenated fosmidomycin analogues. The results showed good inhibition of the enzyme by the phosphonic acids of the chloroacetyl and chloropropyl analogues 2.1 and 2.2 respectively. Molecular modelling of the compounds on E. coli (PDB code: 2EGH) and P. falciparum (PDB code: 3AUA) DXR showed strong binding of the halogenated fosmidomycin analogues while the hybrids in the absence of docked NADPH showed minimum binding to the enzymes.
- Full Text:
- Date Issued: 2012
An investigation of IgE regulation by recombinant soluble IgE receptors and co-receptors in human cell culture models
- Authors: Bowles, Sandra Lyn
- Date: 2010
- Subjects: Immunoglobulin E , Allergy
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:10321 , http://hdl.handle.net/10948/1231 , Immunoglobulin E , Allergy
- Description: Type I hypersensitivities are mediated by the IgE antibody. The effector functions and synthesis of IgE result from interactions with a network of proteins that include a high affinity (FcRIα) and a low affinity (CD23, FcRII) Fc receptor in conjunction with the B lymphocyte receptor, CD21. CD23 is a multifunctional type II transmembrane protein that binds its known ligands through its ectodomain either as a membrane-bound or soluble receptor generated in vivo by specific proteolytic cleavages. IgE production is primarily regulated by interactions between IgE, CD23 and CD21. Despite its importance for development of strategies to limit hypersensitivity, precise information about the molecular interactions remains limited. During this study, I engineered, expressed and purified from bacteria three soluble human CD23 fragments that are normally formed in vivo and shed from the cell surface (1) derCD23, amino acids 156-298 (2) sCD23, amino acids 150-321 and (3) the entire ectodomain, exCD23, amino acids 48-321 to examine the comparative binding of recombinant human CD21 SCR 1-2 and native human IgE to these fragments. Gel filtration HPLC revealed that derCD23 and sCD23 were monomeric whereas exCD23 assembled as a heterogeneous mixture that included trimers and monomers. At the concentrations utilized, CD23 fragments sCD23 and exCD23 bound CD21 with similar affinity, whereas interaction between derCD23 and CD21 was minimal when analyzed by surface plasmon resonance (SPR) spectroscopy. These findings suggest that penultimate “tail” amino acids between 298 and 321 stabilize CD21 attachment, although it cannot be ruled out, the region between Met 150 and Ser 156 may also play a role in binding CD21 SCR 1-2. In contrast, there is a progressive increment in the affinity of soluble fragments (exCD23>sCD23>derCD23) for IgE, upon increasing length of the proximal CD23 “stalk” domain. These findings highlight the differences in both the structural basis and affinity of the three physiological fragments of human CD23 for the ligands CD21 and IgE and underscore the complexity of CD23-mediated regulatory networks. It was found that B-cells only make up ~5% of the PBMC population, and that these cells were able to be activated, via STAT-6 phosphorylation, to enter class switch recombination (CSR) by the addition of switch factors (IL-4 and anti-CD40). Titration experiments dictated that 25 ng/mL of CD23 was the most efficient concentration to up-regulate IgE synthesis in PBMCs; furthermore, soluble CD23 proteins were incubated with PBMCs in the presence and absence of CD21 SCR 1-2 to investigate the effect that these recombinant proteins have on IgE synthesis. Results showed that the influence of recombinant proteins (both CD23 and CD21) on IgE synthesis was slight. It was shown that while derCD23 had no significant effect, monomeric sCD23 down-regulated, and the mixture of monomeric and oligomeric exCD23 up-regulated IgE synthesis. On addition of CD21 SCR 1-2 to the cells switched and treated with soluble CD23, it was found that in both cases for sCD23 and exCD23, IgE synthesis was increased, while for derCD23, there was no noticeable difference in IgE synthesis. This confirmed previous data showing the lack of binding between derCD23 and CD21 SCR 1-2. The exact binding site for CD21 on the CD23 molecule is unknown, and incompletely represented in the NMR and crystal structures. It is thought that CD21 binds to the C-terminal tail section, not present in derCD23. It is therefore likely that only a negative-feedback mechanism operates with derCD23 to regulate IgE synthesis. Further investigation of the binding of CD23 fragments to SCR 5-8 of CD21 and the effect of this on IgE synthesis may lead to a potential therapeutic role for derCD23 in the treatment of allergic disease. Data accumulated in this study suggests that investigating the modulation of oligomeric state and thus the activity of soluble CD23 fragments may be important in the construction of new regulators of IgE synthesis.
- Full Text:
- Date Issued: 2010
The effects of elephants at low densities and after short occupation time on the ecosystems of the Eastern Cape Province, South Africa
- Authors: Parker, Daniel Matthew
- Date: 2008
- Subjects: Elephants -- South Africa -- Eastern Cape Biotic communities -- South Africa -- Eastern Cape Animal populations -- South Africa -- Eastern Cape Plant communities -- South Africa -- Eastern Cape Mammals -- Ecology -- South Africa -- Eastern Cape Animal-plant relationships -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5678 , http://hdl.handle.net/10962/d1005363
- Description: Elephants (Loxodonta africana, Blumenbach) are recognised as keystone herbivores and ecosystem engineers in African ecosystems due to the noticeable impact they have on plant communities. This impact can influence other animal taxa and ecosystem processes, especially within enclosed systems. I investigated the effects of elephants in four vegetation types and the cascade effect on three associated taxa and two ecosystem processes at five sites with elephants and five paired sites without elephants in the Eastern Cape Province, South Africa, between April 2005 and July 2007. In grassland habitats, the physical structure of the vegetation was neither enhanced nor degraded in the presence of elephants. Within the locally important Thicket Biome, elephant browsing transformed relatively homogeneous stands of vegetation into more heterogeneous units. However, although seeds from thicket plant species were recorded in elephant dung, elephants did not promote the viability and germination success of these plants. In addition, elephant foraging on aloes caused declines in their populations. By contrast, the size of bushclumps was not reduced in the presence of elephants in bushclump savanna. The associated insect, bird and mammal communities appeared to benefit from elephant foraging in all vegetation types assessed, whereas vegetation patch dynamics and soil surface processes were neither enhanced nor degraded in the presence of elephants. I conclude that, at current densities, elephants do not (in most cases) negatively affect plant and animal communities or ecosystem processes in enclosed reserves. However, elephants have only been present at each site for a relatively short period and it is likely that their impact will be cumulative, increasing over time in these closed systems. Thus, future research along a continuum of elephant density and time since re-introduction is recommended.
- Full Text:
- Date Issued: 2008
Design, synthesis and evaluation of silver-specific ligands
- Authors: Daubinet, André
- Date: 2001
- Subjects: Ligands Ligands -- Design Ligands -- Analysis Ligands -- Evaluation Silver -- Metallurgy
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4308 , http://hdl.handle.net/10962/d1004966
- Description: Several series of ligands, designed to chelate silver(I) specifically in the presence of base metals, have been synthesised. The ligands include: - dithiodiamide compounds, prepared by the condensation of acetanilide derivatives with 1,2-dibromoethane; propanenitrile and propanoic ester derivatives prepared from pyridine-2-carbaldehyde via the Morita-Baylis-Hillman reaction; and novel malonamide ligands from the reaction of diethyl malonate with a range of primary amines. The malonamide derivatives were prepared under both conventional thermal and microwave-assisted conditions, the latter proving to be highly efficient. The ligands were all characterised using a combination of spectroscopic and, where appropriate, elemental analysis; in one case, the structural assignment was confirmed by single-crystal X-ray analysis. The fragmentation patterns in the electron-impact mass spectra of the malonamide derivatives have been explored using high-resolution and meta-stable peak scanning techniques. Complexes of the malonamide ligands with copper(II) and silver(I) have been synthesised, and examination of these complexes has revealed distinct differences in their co-ordination preferences towards silver(I) and copper(II). Tentative, computer-modelled structures for the complexes have been proposed using the available spectroscopic and elemental analysis data. Computer modelling, at the Molecular Mechanics level, has also been used to assess the capacity of the ligand systems to adopt conformations suitable for the chelation of tetrahedral silver(I). Solvent extraction studies have been undertaken using aqueous metal ion solutions and various organic solvents. The dithiodiamide derivatives typically presented solubility problems, but one of the ligands, N,N´-bis(3-chlorophenyl)-3,6-dithiaoctanediamide, exhibited significant but slow extraction of silver(I) into toluene. The malonamide derivatives, however, proved to be readily soluble in ethyl acetate and, in some cases, exhibited good to excellent selectivity for silver(I) in the presence of the base metals copper and lead. Atomic absorption analysis revealed rapid equilibration times (<15 min) and high extraction efficiencies over a wide pH range (2.78 - 9.0). Metal selectivity has been determined by ICP-MS analysis of the residual silver, copper and lead present in the aqueous phase after 15 min, and one of the ligands, N,N´-bis(2-benzylsulfanylethyl)malonamide, exhibits excellent (≥ 96 %) silver(I) specificity.
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- Date Issued: 2001
An assessment of the monkfish resource of Namibia
- Authors: Maartens, Lima
- Date: 2000
- Subjects: Lophius -- Namibia Lophius
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5231 , http://hdl.handle.net/10962/d1005074
- Description: Two monkfish species are caught off the coast of Namibia. These are Lophius vomerinus Valenciennes (1837) and L. vaillanti Regan (1903). L. vomerinus is the more abundant of the two and contributes approximately 99% to the total Namibian monkfish landings. The Namibian monkfish fishery is one of the largest and most valuable of its kind in the world. Two fishing fleets harvest the resource, viz. the monkfish and sole fleet and the hake directed fleet. In 1998 and 1999, monkfish were also harvested by one gillnet vessel that had an experimental right. In this study, the biology, abundance and exploitation patterns of monkfish were investigated and used as inputs into a stock assessment framework as a contribution towards the development of a management procedure for this species. Ground and sectioned otoliths and sectioned illicia were used to age Lophius vomerinus. The illicia provided more precise estimates of age, and growth was subsequently modelled using these age estimates. Growth in length and weight was best described by the threeparameter specialised von Bertalanffy model as Lt (cm TL) = 72.29 (1-e⁻°·¹⁴⁽t⁺°·³°⁾) and Wt (kg) = 3.96 (1-e⁻°·¹⁸⁽t⁺°·¹°⁾)²·⁹⁹ for males, and Lt (cm TL) = 111.98 (1-e⁻°·°⁸⁽t⁺°·³⁶⁾) and Wt (kg) = 6.92 (1-e⁻°·¹⁸⁽t⁻°·⁶⁴⁾)³·°³ for females. L. vomerinus is a slow-growing species with a life span in excess of 10 years. Reproductive activity was shown to occur throughout the year, with a slight peak during the winter months. The adult sex ratio of fish > 37 cm TL was biased towards females. Length-at-50% sexual maturity was attained at 32.1 cm TL for females and 23.7 cm TL for males. The rate of natural mortality was estimated at 0.15 year⁻¹, while fishing mortality rates during the 1970s, 1980s and 1990s were estimated at 0.01 year⁻¹, 0.04 year⁻¹ and 0.14 year⁻¹, respectively. Gillnetting for monkfish (300 mm stretched mesh) was highly efficient with a moderate bycatch of around 20% during the two years of operation. The main bycatch species were red crab, spider crab, squalid sharks, rays and Cape and Deep-water hake. The mean length of the monkfish caught in gillnets (67 cm TL) was significantly larger than the monkfish landed by the trawlers (38 cm TL) and less than 1% of immature fish were landed. Gillnet catch-per-unit-effort for monkfish fluctuated between 0.03 and 0.67 kg.day⁻¹.50 m net panel⁻¹, with a soak time of between one and sixteen days. More than 50% (by weight) of monkfish landed by monkfish and sole trawlers, consisted of fish below 36 cm TL. There was a significant increase in catches of juvenile monkfish during 1997 and 1998 in comparison to the period 1994 to 1996. Various types of rigid sorting grids were tested to release juvenile monkfish below 32 cm TL. Five grid designs were tested. These included an “Ex-it” grid with horizontal bars spaced at 55 mm, single grids with vertical and horizontal bars spaced at 55 mm and grids with circular openings of 110 and 168 mm in diameter. The most efficient design was the grid with circular openings of 110 cm in diameter, which ensured the escape of 66% of monkfish smaller than 31 cm TL. However, studies need to be undertaken to quantify the survival of released fish and to test the feasibility of using grid sorters on commercial monkfish and sole trawling gear. The monkfish resource was assessed by means of length cohort analyses, the Thompson and Bell predictive model and by way of a deterministic age-structured production modelling approach. The length cohort analysis models were sensitive to the rate of natural mortality and insensitive to changes in the terminal fishing mortality rate. These biases may, however, not be serious provided that estimates of abundance are used to reflect relative changes. Fish ranging between 26 and 59 cm TL are the most heavily exploited. The Thompson and Bell model predicted that the monkfish resource is exploited above MSY -levels and a reduction of approximately 40% in fishing effort would provide a higher yield. Yield-per-recruit ranged between 10 000 and 14 000 tonnes. Results should, however, be treated with caution, as the condition of steady state was not satisfied. The age-structured production model was tuned using trends in catch-per-unit-effort data, estimated by Generalised Linear Modeling, as well as relative abundance indices calculated from hake biomass surveys. The model was found to be sensitive to both the ‘steepness’ parameter h and estimates of natural mortality. The ‘depletion’ level of the monkfish resource is currently estimated to be 49%. Estimated coefficients of variation were high (> 63%) due to the lack of a consistent trend within the abundance indices to tune the model. Overall productivity of the monkfish resource was estimated to be approximately 16%, similar to other southern African demersal resources. Results of the risk analyses suggest that catches in excess of 7 000 tonnes may be unsustainable and that catches of 5 000 or 6 000 tonnes would decrease the risk of stock collapse and possibly lead to a recovery in the stock. Monkfish management strategies were reviewed and these were considered in relation to the results of this study. The following management recommendations were made: to follow the precautionary approach and implement a total allowable catch for monkfish; to implement rigid sorting grids as these would be the most appropriate way in which to reduce catches of juvenile monkfish; to restrict soak time, depth of operation and implement means to reduce ‘ghost fishing’ by gillnetting and finally, to develop a management procedure for Namibian monkfish with the main objective being the sustainable exploitation of the resource.
- Full Text:
- Date Issued: 2000
Zinc inhibition of cell division : its relevance to cancer cells and possible mechanism of action
- Authors: Skeef, Noel Samuel
- Date: 1989
- Subjects: Cell division , Cancer cells -- Growth -- Regulation , Zinc in the body , Zinc -- Physiological effect , Cancer -- Research
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4144 , http://hdl.handle.net/10962/d1016266
- Description: A description of two techniques used extensively in this study namely cell counting with a "cell counting plate" and argentation TLC for the separation of ω -6 -fatty acids is given. Zn supplementation into GM of two malignant (BL-6 and Hep- 350) and a non-malignant (LLC-MK) cell line/s resulted in an increased uptake of Zn by the cells and progressively suppressed proliferation of particularly the malignant cells. Zn chelation by EDTA suppressed in vitro proliferation of all 3 cell line, this effect being more pronounced in the malignant cells. A dietary Zn deficiency resulted in alopecia in mice and both a dietary Zn deficiency and Zn excess reduced growth of BL-6 tumours implanted subcutaneously in mice. Zn supplementation into GM progressively increased the uptake of [1-¹⁴C]-LA by BL-6 and LLC-MK cells but had a very slight though irregular effect on this parameter in the Hep- 350 cells. Zn supplementation also stimulated desaturase activity in the BL-6 cells. These results suggested that there are select cell lines whose Δ⁶-desaturase activity responds positively to Zn supplementation (e.g. the BL-6 cells). Delta-6-desaturase activity was also assayed in microsome preparations from different tissues. No enzyme activity was detected in the microsomes prepared from the BL-6 tumours. There was no significant effect with the addition of Zn or EDTA, on Δ⁶-desaturase activity of the regenerating liver microsomes. In the resting liver microsomes this enzyme activity was reduced only when EDTA and Zn were added together and when EDTA was added to the reaction medium as well as to the microsome preparations 2 hr before the enzyme activity assay was initiated. The results of these experiments suggested that the Δ⁶-desaturase enzyme in the microsome preparations may have had an adequate amount of Zn with further additions having no stimulatory effect on the enzyme. Two independent mechanisms of control of cell proliferation by low and high Zn are suggested to operate.
- Full Text:
- Date Issued: 1989