Charles K.S. Moy

Development and characteristics of polymer monoliths for advanced LC bioscreening applications: A review

Biomedical research advances over the past two decades in bioseparation science and engineering have led to the development of new adsorbent systems called monoliths, mostly as stationary supports for liquid chromatography (LC) applications. They are acknowledged to offer better mass transfer hydrodynamics than their particulate counterparts. Also, their architectural and morphological traits can be tailored in situ to meet the hydrodynamic size of molecules which include proteins, pDNA, cells and viral targets. This has enabled their development for a plethora of enhanced bioscreening applications including biosensing, biomolecular purification, concentration and separation, achieved through the introduction of specific functional moieties or ligands (such as triethylamine, N,N-dimethyl-N-dodecylamine, antibodies, enzymes and aptamers) into the molecular architecture of monoliths. Notwithstanding, the application of monoliths presents major material and bioprocess challenges. The relationship between in-process polymerisation characteristics and the physicochemical properties of monolith is critical to optimise chromatographic performance. There is also a need to develop theoretical models for non-invasive analyses and predictions. This review article therefore discusses in-process analytical conditions, functionalisation chemistries and ligands relevant to establish the characteristics of monoliths in order to facilitate a wide range of enhanced bioscreening applications. It gives emphasis to the development of functional polymethacrylate monoliths for microfluidic and preparative scale bio-applications.

Macroscopic electrical field distribution and field-induced surface stresses of needle-shaped field emitters.

One major concern since the development of the field ion microscope is the mechanical strength of the specimens. The macroscopic shape of the imaging tip greatly influences field-induced stresses and there is merit in further study of this phenomenon from a classical perspective. Understanding the geometrical, as opposed to localized electronic, factors that affect the stress might improve the quality and success rate of atom probe experiments. This study uses macroscopic electrostatic principles and finite element modelling to investigate field-induced stresses in relation to the shape of the tip. Three two-dimensional idealized models are considered, namely hyperbolic, parabolic and sphere-on-orthogonal-cone; the shapes of which are compared to experimental tips prepared by electro-polishing. Three dimensional morphologies of both a nano-porous and single-crystal aluminium tip are measured using electron tomography to quantitatively test the assumption of cylindrical symmetry for electro-polished tips. The porous tip was prepared and studied to demonstrate a fragile specimen for which such finite element studies could determine potential mechanical failure, prior to any exhaustive atom probe investigation.

Deploying aptameric sensing technology for rapid pandemic monitoring

Abstract The genome of virulent strains may possess the ability to mutate by means of antigenic shift and/or antigenic drift as well as being resistant to antibiotics with time. The outbreak and spread of these virulent diseases including avian influenza (H1N1), severe acute respiratory syndrome (SARS-Corona virus), cholera (Vibrio cholera), tuberculosis (Mycobacterium tuberculosis), Ebola hemorrhagic fever (Ebola Virus) and AIDS (HIV-1) necessitate urgent attention to develop diagnostic protocols and assays for rapid detection and screening. Rapid and accurate detection of first cases with certainty will contribute significantly in preventing disease transmission and escalation to pandemic levels. As a result, there is a need to develop technologies that can meet the heavy demand of an all-embedded, inexpensive, specific and fast biosensing for the detection and screening of pathogens in active or latent forms to offer quick diagnosis and early treatments in order to avoid disease aggravation and unnecessary late treatment costs. Nucleic acid aptamers are short, single-stranded RNA or DNA sequences that can selectively bind to specific cellular and biomolecular targets. Aptamers, as new-age bioaffinity probes, have the necessary biophysical characteristics for improved pathogen detection. This article seeks to review global pandemic situations in relation to advances in pathogen detection systems. It particularly discusses aptameric biosensing and establishes application opportunities for effective pandemic monitoring. Insights into the application of continuous polymeric supports as the synthetic base for aptamer coupling to provide the needed convective mass transport for rapid screening is also presented.

Next-generation sequencing showing potential leachate influence on bacterial communities around a landfill in China

The impact of contaminated leachate on groundwater from landfills is well known, but the specific effects on bacterial consortia are less well-studied. Bacterial communities in a landfill and an urban site located in Suzhou, China, were studied using Illumina high-throughput sequencing. A total of 153 944 good-quality reads were produced and sequences assigned to 6388 operational taxonomic units. Bacterial consortia consisted of up to 16 phyla, including Proteobacteria (31.9%-94.9% at landfill, 25.1%-43.3% at urban sites), Actinobacteria (0%-28.7% at landfill, 9.9%-34.3% at urban sites), Bacteroidetes (1.4%-25.6% at landfill, 5.6%-7.8% at urban sites), Chloroflexi (0.4%-26.5% at urban sites only), and unclassified bacteria. Pseudomonas was the dominant (67%-93%) genus in landfill leachate. Arsenic concentrations in landfill raw leachate (RL) (1.11 × 103 μg/L) and fresh leachate (FL2) (1.78 × 103 μg/L) and mercury concentrations in RL (10.9 μg/L) and FL2 (7.37 μg/L) exceeded Chinese State Environmental Protection Administration standards for leachate in landfills. The Shannon diversity index and Chao1 richness estimate showed RL and FL2 lacked richness and diversity when compared with other samples. This is consistent with stresses imposed by elevated arsenic and mercury and has implications for ecological site remediation by bioremediation or natural attenuation.

Stimulation of Indigenous Carbonate Precipitating Bacteria for Ground Improvement

Calcite minerals are precipitated in soil through biomineralisation which can be either organic or inorganic in nature. Biomineralisation can be employed to improve ground conditions in its natural state. Usually, studies of applied biomineralisation are highly interdisciplinary involving expertise from engineers, chemists and microbiologists. In this paper, we study the potential of biomineralisation from indigenous bacteria present in soil. The soil samples were collected from a high permeable zone and the bacteria that inhabit the soil were stimulated at a temperature of 15°C. A cementation solution consisting of 500mM calcium chloride, urea and nutrient broth at a pH of 7.5 was added to the soil samples. Inorganic precipitation was found to be dominant and was more efficient when compared to organic precipitation. Carbonate precipitation data indicated that inorganic precipitation were 1.37 times better at carbonate formation in comparison to organic precipitation. Scanning Electron Microscopy analysis identified cementation bonds formed between soil particles. It was deducted that organic precipitation is dependent on temperature, and may take an extended time at such low temperature. The preliminary data presented in this paper suggests that the implementation of biomineralisation with in-situ microbes is promising but requires further laboratory and field investigation before being considered for engineering application.

MICP and Advances towards Eco-Friendly and Economical Applications

Biomineralization is a natural process aided by living organisms. Due to its applicability in ground improvement and bioremediation, Microbially Induced Calcite Precipitation (MICP) is an interdisciplinary field of study combining engineering, chemistry and microbiology. Bioremediation has been applied widely for contamination containment or removal, in this case it will be containment. MICP can also be applied to improve the efficiency of insitu bioremediation. Urease is an enzyme which can facilitate increased calcite precipitation. However the production of urease by bacteria and thus the resulting carbonate precipitation are inhibited by environmental factors including calcium concentration, bacterial concentration, pH and temperature. Under good conditions MICP can be used for heavy metal and radionuclide immobilization. However technologies such as bioconsolidation and biocementation require improvement such as time and cost. This paper highlights the application of MICP in addition to suggested improvements to make it more eco-friendly and sustainable.

Chromatographic characterisation of aptamer-modified poly(EDMA-co-GMA) monolithic disk format for protein binding and separation

ABSTRACT The introduction of aptameric ligands onto disk-monolithic adsorbent, representing a unique strategy for convective isolation of target molecules with high specificity and selectivity, is investigated for the first time. Experimental results showed that the disk monolith possessed a good permeability of 1.67 ± 0.05 × 10–14 m2 (RSD = 3.2%). The aptameric ligand density for the aptamer-modified disk monolith was 480 pmol/uL. Chromatographic analysis of the aptamer disk-monolith efficiency showed an optimum linear velocity of 126 cm/min (≈0.25 mL/min) at room temperatures 25 ± 2°C. The theoretical number of plates corresponding to the optimum linear velocity was 128.2 with an height equivalent to the theoretical plate of 0.022 mm. The disk aptamer-immobilised monolithic system demonstrated good selectivity and isolation of thrombin from non-targets.

Effects of preparation methods on inherent fabric anisotropy and packing density of reconstituted sand

Abstract This paper investigates the effects of different preparation methods (dry tamping, moist tamping, dry pluviation and wet pluviation) on inherent fabric anisotropy of reconstituted sand samples in the laboratory. An image analysis approach was used to quantify the inherent fabric anisotropy in terms of the vector magnitude based on a second-order fabric tensor. In addition, the effects of reconstituted soil sample preparation methods on the void ratio and density index were investigated. This study shows that the specimens prepared by pluviation methods possess more pronounced inherent fabric anisotropy than those by tamping methods. Among all the methods considered in this investigation, the dry pluviation method provided the highest degree of inherent fabric anisotropy. It is shown that the mass-flow and drop height does not effectively influence the packing density. Also, it has been found that no significant effect on void ratio is observed while using different tamping methods, which may be attributed to the fact that the sand used in this study is less well graded. In addition, it is found that the increase in the height of tamper leads to an increase in packing density or a decrease in relative density, with an approximately linear relationship.

Biomineralisation performance of bacteria isolated from a landfill in China

We report an investigation of microbially induced carbonate precipitation by seven indigenous bacteria isolated from a landfill in China. Bacterial strains were cultured in a medium supplemented with 25 mmol/L calcium chloride and 333 mmol/L urea. The experiments were carried out at 30 °C for 7 days with agitation by a shaking table at 130 r/min. Scanning electron microscopic and X-ray diffraction analyses showed variations in calcium carbonate polymorphs and mineral composition induced by all bacterial strains. The amount of carbonate precipitation was quantified by titration. The amount of carbonate precipitated in the medium varied among isolates, with the lowest being Bacillus aerius rawirorabr15 (LC092833) precipitating around 1.5 times more carbonate per unit volume than the abiotic (blank) solution. Pseudomonas nitroreducens szh_asesj15 (LC090854) was found to be the most efficient, precipitating 3.2 times more carbonate than the abiotic solution. Our results indicate that bacterial carbonate precipitation occurred through ureolysis and suggest that variations in carbonate crystal polymorphs and rates of precipitation were driven by strain-specific differences in urease expression and response to the alkaline environment. These results and the method applied provide benchmarking and screening data for assessing the bioremediation potential of indigenous bacteria for containment of contaminants in landfills.

Structural Performance of a Precast Beam to Column Connection Strengthened with FRP under Monotonic Loading

Over the years, precast concrete construction has been employed in many parts of the world mainly because it is less labour intensive and rapid. However, its connection remains in many cases an issue that requires careful consideration at both the design and construction stages. More precisely, the ductility and rotational stiffness are often challenging factors that need to be considered. In that perspective, this work focuses on finding the improvement of a precast beam-to-column connection with strengthening from fibre reinforced polymer (FRP). High strength basalt fibre fabric was used in this particular study. The study revealed a net improvement in the connection’s rotational stiffness and ductility. It was also found that the addition of a steel plate on top of the FRP contributed to maintaining the strength of the connection under late plastic deformation.