新加坡

a. Purpose of research- Since the discovery of genetic codes and the dogma of 64 codons coding for 21 amino acids, scientists worldwide have been interested to know the reason(s) behind this unique number ratio (64:21). This ratio indicates certain form of inefficiency in the replication of amino acids. Such inefficiency can be explained through symmetries in the condons coding for the same amino acids. In the light of that, my project looks for patterns in the properties of amino acids and symmetries in the codons combinations. Using these analysis findings, I invented a three dimensional periodic table for the codons and amino acids that has a points to layers ratio of 64:21. b. Procedures- To get started with the project, I searched for relevant information in books and the Internet. After locating the relevant materials, I began my analysis by looking for non-random patterns in the correlation between codons and the respective amino acids they code for. At the same time, I try to look for symmetries in the codon distributions and suggest new and innovative models for a periodic table of codon combinations. I have come out with mainly a new model, with its own unique ideas and concepts behind it. Finally, I will try to match a property of the amino acids to the positions of the codons such that the table shows a gradual change in property of the amino acids, together with the symmetries. This will effectively explain the unique codons to amino acids ratio and lead to discovery of possible amino acids. c. Data- This research is primarily conducted based on the conventional 2D periodic table and no experimental data is collected. After much analysis, I have come up with the 3-sided triangular pyramid model. This model is inspired by the ratio of 64 codons coding for 21 amino acids, which can be easily approximated to 3:1. It is made up of a triangular pyramid that is three-faced, with the bottom side unutilized. As a triangular structure, each layer has dimensions in the multiples of 3. Layer 1 consists of 1 point, layer 2 with 3 points, layer 3 with 6 points and so on… until layer 7 with 18 points, having a total of 64 points. This 64 points to 21 layers ratio is consistent with the codons to amino acids ratio! d. Conclusions- The unique 64:21 ratio suggest certain form of inefficiency in the replication of amino acids. This may be explained through symmetries in condons coding for the same amino acids. A general 3:1 ratio can be approximated and this suggests a high possibility for the existence of a three-sided symmetry in codon combinations. Thus, this idea of a three-sided symmetry gives rise to my 3-sided triangular pyramid model. This new model of a 3-dimensional periodic table for codon combinations would be useful in explaining such a unique 64:21 ratio and serves to provide a basis for better understanding of the relationship between codons and amino acids. This new model may also lead to the discovery of currently unknown amino acids.

In this work, techniques to explore the capabilities of multi-walled carbon nanotubes\r (MWNTs) in sorting nanoparticles (NPs) were presented. A droplet of a solution comprising of quantum dots (QDs) with various sizes was deposited on an aligned array of intertwined MWNTs. Photoluminescence (PL) and fluorescence microscopy (FM) revealed that MWNTs were effective nano-sieves that could effectively sort out QDs with a size difference of ~ 2.1 nm.\r Cadmium Selenide/Zinc Sulfide (CdSe/ZnS)core-shell QDs and Cadmium Sulfide (CdS) QDs were used to explore whether chemical properties of NPs affect the sieving capability of MWNTs. Further investigation on the effects of micro-patterning on the sieving ability of MWNTs was also carried out.PL and FM results suggested that micro-patterning could aid in separation of QDs and thus improve sieving capability of MWNTs. With the above findings, QDs emitting different colors as a result of size difference could efficiently be assembled onto the MWNTs en route to three-dimensional architectures with controlled assembly of NPs.\r Together with controlled laser power to remove desired amounts of QDs decorated MWNTs, a multi-colored display could be achieved. Further experiments were also carried out to determine the feasibility of introducing MWNTs as filters for NPs. Dilute solutions containing NPs such as gold colloid was run through these MWNTs filters by gravity. Field emission scanning electron microscope (FESEM) images of the samples showed that MWNTs were successful in trapping the nanoparticles. Explorations into the length dependent effect of using MWNTs as filters, suggested that 300μm MWNTs are better nano-sieves compared to 50μm MWNTs.

The physiochemical properties of fullerenes have aroused wide interest, such as its ability to accept and lose electrons and relatively high reactivity that permit various modes of structural modifications. However, obstacles to further research include its complete lack of solubility in water and low processability.\r This project investigated the morphology and microstructure of a fullerene-grafted polymer as a potential candidate for better and novel systems for drug delivery. In this research, hydrophilic functionalities were introduced to the C60 fullerene by chemical modifications, through the attachment of poly(acrylic acid) (PAA) chain. The objective was to investigate the dynamics and the self-assembly properties of this polymer in aqueous solutions, and the knowledge gained would enhance the development of such system for potential applications in drug delivery and nanotechnology.

NO donors are an emerging class of pharmaceutical compounds, with many important functions in the cardiovascular, nervous and immune systems. With great therapeutic potential, the development of new NO donor compounds would be of great medicinal value, potentially opening a whole class of drugs to be used to treat various ailments. This project studies a specific class of compounds, substituted cyclic oxime ethers, which have proven to be useful intermediates in fields such as medicine and biochemistry. The cyclic structure along with a determinable substitutable group at the C3 position is highly valuable, as it allows the oxime ether to act as a convenient precursor for a variety of useful products, playing key components in many drugs. And with a substituted nitrate group, which is an O-nucleophile, the oxime ether has the potential to become an NO-donor, and hence become a possible intermediate in a wide array of NO donor drugs. Co(NO)3 was used in the synthesis of the cyclic oxime ether, directly from a phenyl substituted bis(oxy)enamine intermediate, producing an entirely new compound: α-hydroxyoxime nitrates, the oxime ether being substituted with a nitrate group. This new reaction of the synthesis of α-hydroxyoxime nitrates was further studied for optimization purposes, in order to open a new class of NO donor precursors. In addition, other nucleophiles were also explored in this class of reactions, forming important bonds such as C-N and C-S bonds, with key structures for other types of synthesis intermediates and precursors. Different metal nitrates, or various other nucleophiles in place of the nitrates, were used in reaction with bis(oxy)enamine, and the yield and structure of the final products were determined by NMR spectra. Successful optimization of the synthesis of α-hydroxyoxime nitrates has been achieved, where the conditions for optimum synthesis involve using Cr(NO3)3•9H2O which achieved a high yield of 76%, dissolved in THF with the bis(oxy)enamine starting compound. It has been determined that the metal in the salt affects the reaction pathway, as the nature of the metal cation affects its efficiency to cleave the N-O bond in the starting compound (with d-block elements being the best performing), and H+ ions can promote the reaction as well. Also, the reaction proceeds with different types of bis(oxy)enamines, meaning the substrate scope can be expanded to give a variety of products. The reaction can also proceed to form other products with different nucleophiles other than the nitrate group, where the C-N and C-S bonds were successfully formed in the reactions from bis(oxy)enamine to oxime ether. Thus, this class of reaction in converting the bis(oxy)enamine to a cyclic oxime ether has potentially opened a new class of NO donor compounds, and further possesses the potential to form a wide variety of products to be used in other important synthesis procedures.

Elevation of intracellular calcium secondary to increased calcium influx along with increased gliosis are implicated in the pathogenesis of focal ischemic stroke. In astrocytes, which play a major role in maintaining homeostasis in brain ischemia, the identities of the ion channels responsible for increased calcium influx during ischemia is relatively unknown although several Ca2+-permeable transient receptor potential (TRP) channels have been identified to have contributing roles. The transient receptor potential vanilloid 5 (TRPV5) channel is a Ca2+-permeable cationic channel expressed primarily in kidney epithelial cells and at low levels in the brain, but the exact localization and role this channel plays in the brain has not been explored. To investigate the possible role TRPV5 plays in astrocytic calcium influx in ischemia, we examined the functional expression of TRPV5 in astrocytes subjected to hypoxia-ischemia in vitro and in rat models of ischemic stroke in vivo. We hypothesize that TRPV5 contributes to increased calcium influx in ischemia. By treating astrocytes with culture conditions without glucose and with low oxygen levels, we found that TRPV5 is upregulated with increasing durations of simulated hypoxia-ischemia in vitro. Similarly, rat models of ischemic stroke with middle cerebral artery occlusion also show TRPV5 upregulation in reactive astrocytes, suggesting a possible role of TRPV5 in reactive gliosis in vivo. Microfluorimetric intracellular calcium imaging using Fura-2 on primary cultured astrocytes show a voltage-independent increase in astrocytic calcium influx after hypoxia-ischemia in vitro that is selective for extracellular Ca2+ concentration and is reduced by inhibition of TRPV5 with ruthenium red. Electrophysiology measurements using the whole-cell patch clamp technique on primary cultured astrocytes reveal a non-selective cation current similar to that of TRPV5 that is inhibited by Mg2+, another inhibitor of TRPV5. Preliminary results on astrocyte cell viability during hypoxia-ischemia with TRPV5 inhibition by ruthenium red also suggest that inhibition of TRPV5 could enhance astrocyte survival and reactive gliosis in vitro, indicating a beneficial role in blocking non-selective Ca2+ entry via TRPV5 into astrocytes. Since TRPV5 is highly selective for Ca2+ and an important channel for Ca2+ absorption in various epithelial cells, TRPV5 upregulation may contribute significantly to elevated Ca2+ influx in astrocytes in hypoxia-ischemia. Also, Ca2+ influx has been demonstrated to play a crucial role in reactive gliosis, further suggesting that TRPV5 upregulation is involved in reactive gliosis. We propose that TRPV5 is involved in ischemia-induced calcium influx in astrocytes, and might participate in the pathogenesis of focal ischemic stroke.

Lipocalin 2 (LCN2), a 25-kDa secreted protein that belongs to the lipocalin family, is known to bind to a class of bacterial Fe-binding molecules known as siderophores. Iron is essential for bacterial growth. To obtain iron from host cells, bacteria produce siderophores, such as Enterochelin (Ent), to bind and transport host iron into the bacterial cell. In response, the host produces LCN2 to bind the iron-laden enterochelin, forming the tricomplex, LCN2: Ent: Fe3+. This inhibits bacterial growth as iron has been sequestered by LCN2. Devireddy et.al. 2005, proposed the binding of the tricomplex, LCN2: Ent: Fe3+ with the LCN2 receptor (LCN2R). This resulted in the internalisation of the complex, releasing the bound iron into the cell. The increase of intracellular iron was reported to cause cell mortality. Recent publications postulated 2,5-dihydroxybenzoic acid (2,5-DHBA) to be an endogenous mammalian siderophore homologue in mouse in vivo and in vitro studies, which could sequester LCN2 and iron. High iron concentrations in the brain have been consistently observed in Alzheimer's disease and Parkinson's disease. Accumulation of intracellular iron is known to be toxic to neurons, resulting in neurodegeneration. Hence, this study aims to determine the role of 2,5-DHBA as the mammalian siderophore in a cell culture model of neurodegeneration. We hypothesise that addition of 2,5-DHBA to cells exposed to LCN2 will result in increased iron uptake into neuronal cells, reducing cell viability. SH-SY5Y (human neuroblastoma) cell line was used in our study. To determine if SH-SY5Y is a suitable cell line, endogenous levels of LCN2 and LCN2R mRNA and protein expression were determined using reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot analysis respectively. Preliminary results showed presence of both the LCN2R mRNA and protein but absence of LCN2 mRNA. This could be due to the low expression of LCN2 when not exposed to stress. Hence, to simulate conditions of neurodegeneration (by inducing high expression of LCN2), SH-SY5Y was treated with Kainic Acid (KA). After KA, LCN2 mRNA and protein expression levels will be detected again. With the successful upregulation of LCN2 gene expression, SH-SY5Y will be treated with 2,5-DHBA with KA treatment to determine cell viability using the MTS cell proliferation assay. A decreased cell viability or increased expression of pro-apoptotic genes would support the function of 2,5-DHBA as a mammalian siderophore in the brain. Furthermore, KA treatment can also be applied to microglial or astrocyte cell lines, which are known to secrete high levels of LCN2 when treated with KA. Co-culturing these cells with SH-SY5Y can allow us to study the downstream effects of secreted LCN2 from glial cells binding to the LCN2R receptors on SH-SY5Y neuronal cells. This study will help to further understanding of the relationship between 2,5-DHBA and cellular iron transport. If 2,5-DHBA is able to bind LCN2 and iron to increase intracellular iron levels in the neuronal cells, the formation of the tricomplex, LCN2: 2,5-DHBA: Fe3+, could be targeted for therapeutic interventions in neurodegenerative diseases by reducing intracellular iron levels to help ameliorate the progression of neurodegenerative diseases.

The objective of the project is to develop a novel biomimetic membrane and/or a scaffold for the said membrane. The approach of the project is to use animal skin from the domesticated pig or fish as a scaffold material for the adherence and growth of human skin fibroblasts to create a biomimetic membrane that can be used in medical applications as an alternative to today’s gold standards of Xenograft, Allograft and Autograft procedures. The biomimetic skin membrane can be used to treat victims of burns or scarring with a natural material that would be eliminated via natural bodily functions while eliminating the side effects and drawbacks such as scarring, secondary infections and tissue damage resulting from the current gold standard graft procedures on donor sites. Pig and fish skins were treated with ethanol and dehydrated followed by perfusion with Phosphate buffer solution and Cell culture media. Human skin fibroblasts (NF3 cells) were seeded on the animal skin scaffold. The human skin fibroblasts were then observed to determine their morphology and membrane formation properties. It was observed that the human skin fibroblasts were able to adhere to the non-human skin scaffolding and proliferate. More research is needed to determine their viability as a biomimetic membrane.

The aim of our research is to produce superhydrophobic coatings on both glass and cloth substrates in order to achieve high contact angles and low sliding angles for self-cleaning. In addition, we aim to modify these coatings to be as transparent as possible so as not to interfere with the aesthetics of the objects which will be coated. To achieve this goal, we synthesised a solution using 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (a type of FAS), silica nanoparticles (SiO2), tetraethyl orthosilicate (TEOS), (3-glycidyloxypropyl) trimethoxysilane (Glymo) and deionised water. Using the convenient sol-gel method, coatings of 20% and 30% by weight of FAS-SiO2 nanoparticles were prepared on glass and cotton substrates. It was found that coatings containing 30% by weight of FAS-modified SiO2 nanoparticles on glass slide produced coatings with water contact angle as high as 162.8° and sliding angle as low as 4°. It can also be seen that for glass substrates, the hydrophobicity increased with an increase in percentage of FAS-modified SiO2 nanoparticles. Although the highest percentage transmittance was about 30%, texts and pictures beneath the coated glass slides were clearly readable. The cotton substrates also exhibited excellent hydrophobicity, with a water contact angle of 150° and sliding angle of 22°. Furthermore, the substrates showed good retention of colour and durability after simulated washing and 72 hours of ultraviolet (UV) weathering chamber test. These results show that the effects of washing and UV on the important properties of the cloth were insignificant.

1 Purpose of the Research Nanocrystal thin films exhibit many useful properties, including electrochromicity and superconductivity. When synthesised via Molecular Beam Epitaxy (MBE), selection of substrate, specifically knowledge of crystal orientation, is critical. Reflection High Energy Electron Diffraction (RHEED) is an in situ crystal characterisation method highly compatible with MBE. This study explores a new method of RHEED analysis to determine crystal orientation. 2 Procedure/Theoretical Framework RHEED characterization is the incidence of a beam of high-energy electrons at a low angle with respect to the sample surface. Electrons diffract, and interfere to form patterns on the detector. Traditionally, studies of RHEED analyse one static image as a representation of the surface structure, or observations of RHEED patterns over time. The approach to RHEED analysis in this study exploits changes in RHEED patterns given a rotating substrate. Having specific rotational symmetries along different axes, crystal structures can be differentiated by determining rotational symmetry through RHEED. Electrons scatter upon incidence with crystal planes within the crystal to form Kikuchi lines on the RHEED detector (Fig. 2). The orientation of crystal with respect to incident electron beam affects the Kikuchi line patterns. If the crystal is rotated, crystal planes change orientation, and electrons would diffract from crystal planes in different directions. As such, as the crystal is rotated, the Kikuchi lines move. When the degree of rotation of the crystal corresponds to the rotational symmetry of the crystal (Fig. 1), the Kikuchi lines return to their original position. As crystals with different crystal plane orientations exhibit different orders of symmetries, analyzing the Kikuchi line patterns of the crystal at different degrees of rotation can reveal the rotational symmetry and consequently crystal plane orientation of a crystal. 3 Data/Experimental Testing In order to assess the practical viability of the proposed method, experiments were conducted on SrTiO3 (001), (110), and (111). SrTiO3 exists as a typical perovskite structure (Fig. 3), often used in the synthesis of superconductors via MBE. 3.1 Methodology RHEED images of each sample were taken at 0◦, 60◦, 90◦ and 180◦. Curves were fit to each Kikuchi line observed in the image (Fig. 4). These Kikuchi line approximations are compared by superimposing the curves traced and qualitatively assessing the degree of similarity between the Kikuchi lines of 2 images, to verify the order of symmetry and crystal orientation of the crystal. In the images of the superimposed Kikuchi lines illustrated in Fig. 5, there is similarity between the Kikuchi lines when only when the sample has been rotated by an angle corresponding its degree of symmetry. 4 Conclusions This study offers a method to determine the crystal orientation of thin film through determining the degree of rotational symmetry of the sample, by observation of Kikuchi lines in the RHEED pattern as the sample is rotated. Experimental data was analyzed qualitatively to verify the viability of this theoretical method in practice. This method could be extended to analyze the symmetry of other crystal structures. As it does not require information on the machine settings or usage of complex functions to produce a reliable output, this method is fast and straightforward, opening doors to more streamlined RHEED analysis.

Dronedarone is an anti-arrhythmic drug approved in 2009 for paroxysmal and persistent atrial fibrillation. It is less toxic than its predecessor Amiodarone as it does not cause systemic toxicity but has the same pharmacological activity. However the administration of dronedarone to permanent AF and heart failure patients leads to increased risk of stroke and cardiac death. The exact mechanism of the toxicity is currently unknown. Extrahepatic Cytochrome P450 enzymes play a dominant role in organ-specific drug metabolism and toxicity. Cytochrome P450 2J2 (CYP2J2) enzyme, a predominant enzyme found in human cardiac myocytes, metabolizes endogenous arachidonic acid (AA) into epoxyeicosatrienoic acids (EETs) which play an important role in maintaining normal cardiac physiology. Inhibition of CYP2J2 and perturbation of AA metabolic pathway could result in exacerbation of cardiac failure. This research aims to find out whether dronedarone inhibits CYP2J2 in a suitable cell model (H9C2) using astemizole as a probe substrate. Our in-house studies using recombinant CYP2J2 enzyme have shown that dronedarone potently inhibits CYP2J2. Rat myoblast cells (H9C2) will be seeded in 12-well plate and differentiated for 4 days. The cells will be then treated with different concentrations of astemizole and incubated for 24 h. The cells will then be harvested, lysed, and the cell lysate will be analyzed using liquid chromatography-mass spectrometry (LCMS). Using multi-reaction monitoring (MRM) on the LCMS, astemizole concentration as well as its CYP2J2-specific metabolite O-desmethylastemizole concentrations will be measured. The presence of O-desmethylastemizole confirms the metabolism of astemizole by CYP2J2 in H9C2 cells. By plotting a Michaelis-Menten kinetics curve, we will be able to determine the Michaelis constant (KM) and maximum rate of reaction (Vmax). H9C2 cells will be then treated with fixed concentration of astemizole while varying the dronedarone concentration. A decrease in metabolite O-desmethylastemizole conce ntration, indicates inhibition of CYP2J2 metabolism by dronedarone. Using this data, Lineweaver-Burke graph will be plotted, to determine the mode and potency of the inhibition. Our preliminary studies showed that the KM value was 2.7μM. This study will be useful in understanding if dronedarone inhibits CYP2J2 which may lead to clinically significant drug-drug interactions, one of the dangers of polypharmacy. Finally this study will shed a new light on the mechanisms for dronedarone mediated cardiac failure exacerbation.