Facilitating teacher leadership in Kazakhstan

This is an action-based study that was conducted in four schools in Kazakhstan with the purpose to facilitate teacher leadership for sustainable improvement of practice and enhancing the teacher’s role in education reform in Kazakhstan. By drawing on a non-positional approach to teacher leadership, this study sought bottom-up approaches to educational reform, school leadership and professional development in schools in Kazakhstan. The nine-month intervention programme, which was called the Teacher Leadership for Learning and Collaboration, introduced strategies and created conditions for teachers to lead educational improvement at classroom, school and system level. This has implications for a wider societal development. With the dissolution of Soviet Union, Kazakhstan has been undergoing transition for the last few decades. In the new era, the country is seeking the revival of its national identity, inclusion of its citizens and economic competitiveness, wherein educational improvement has become the country’s top priority. The outcomes of this study indicate that educational improvement requires building local capacity by empowering teachers to lead learning and innovation within and outside their schools. Such a grassroots movement requires systemic and systematic facilitation in schools in Kazakhstan. Ongoing practice-oriented critical reflection, focused action, horizontal communication and school networking can enable teachers and school leadership team members to develop their understanding and foster participatory practices within schools. The study employed a critical participatory action research approach that enabled the building of context-sensitive knowledge and included the voices and reflections of the participants involved in this research. Data were collected through multiple research tools, analysed both during and after the intervention process and presented in the form of a critical narrative to depict the nuances of local reality. The outcomes of this study suggest that teachers can innovate, engage in creative pedagogical practice and lead educational improvement, which requires developing strategies and involving all stakeholders in facilitating teacher leadership.'Talap' scholarship, Nazarbayev University, Kazakhsta

Efficient Separation and Sensitive Detection of Listeria Monocytogenes Using Magnetic Nanoparticles, Microfluidics and Interdigitated Microelectrode Based Impedance Immunosensor

Listeria monocytogenes continues to be a major foodborne pathogen that causes food poisoning and sometimes death in immunosuppressed people and abortion in pregnant women. Nanoparticles have recently drawn attentions for use in immunomagnetic separation techniques due to their greater surface area/volume ratio and better stability against sedimentation in the absence of a magnetic field. Interdigitated microelectrodes and microfluidics make material transfer more efficient and biological/chemical interaction between the surface and solution phase much quicker. Magnetic nanoparticles (Fe3O4) with a 30 nm diameter were functionalized with rabbit anti-L. monocytogenes antibodies via biotin-streptavidin bonds and then amalgamated with target bacterial cells to capture them during a 2 h immunoreaction. A magnetic field was applied to capture the nanoparticle-L. monocytogenes complexes and the supernatant was removed. After a washing step, L. monocytogenes was separated from a food sample and could be ready for detection by a microfluidics and interdigitated microelectrode based impedance biosensor. Capture and separation efficiency of 75% was obtained with the magnetic nanoparticles for L. monocytogenes in phosphate buffered saline (PBS) solution. When combined with the microfluidics and interdigitated microelectrode, the lower detection limits of L. monocytogenes in pure culture and food matrices were 10^3 and 10^4 CFU/ml, respectively, which were equivalent to several bacterial cells in 34.6 nl volume of a sample injected into the microfluidic chamber. A linear correlation was found between the impedance change and target bacteria in a range of 10^3-10^7 CFU/ml. Equivalent circuit analysis indicated that the impedance change was mainly due to the decrease in medium resistance when L. monocytogenes cells attached to the magnetic nanoparticle-antibody conjugates in mannitol solution. The separation and detection of L. monocytogenes were not affected by presence of other foodborne bacteria. A specific, sensitive, and reproducible method using the microfluidics and interdigitated microelectrode based impedance immunosensor in couple with antibody conjugated magnetic nanoparticles was able to detect L. monocytogenes as low as 10^3 CFU/ml in 3 h

CFD Modeling of Chamber Filling in a Micro- Biosensor for Protein Detection

Tuberculosis (TB) remains one of the main causes of human death around the globe. The mortality rate for patients infected with active TB goes beyond 50% when not diagnosed. Rapid and accurate diagnostics coupled with further prompt treatment of the disease is the cornerstone for controlling TB outbreaks. To reduce this burden, the existing gap between detection and treatment must be addressed, and dedicated diagnostic tools such as biosensors should be developed. A biosensor is a sensing micro-device that consists of a biological sensing element and a transducer part to produce signals in proportion to quantitative information about the binding event. The micro-biosensor cell considered in this investigation is designed to operate based on aptamers as recognition elements against Mycobacterium tuberculosis secreted protein MPT64, combined in a microfluidic-chamber with inlet and outlet connections. The microfluidic cell is a miniaturized platform with valuable advantages such as low cost of analysis with low reagent consumption, reduced sample volume, and shortened processing time with enhanced analytical capability. The main purpose of this study is to assess the flooding characteristics of the encapsulated microfluidic cell of an existing micro-biosensor using Computational Fluid Dynamics (CFD) techniques. The main challenge in the design of the microfluidic cell lies in the extraction of entrained air bubbles, which may remain after the filling process is completed, dramatically affecting the performance of the sensing element. In this work, a CFD model was developed on the platform ANSYS-CFX using the finite volume method to discretize the domain and solving the Navier–Stokes equations for both air and water in a Eulerian framework. Second-order space discretization scheme and second-order Euler Backward time discretization were used in the numerical treatment of the equations. For a given inlet–outlet diameter and dimensions of an in-house built cell chamber, different inlet liquid flow rates were explored to determine an appropriate flow condition to guarantee an effective venting of the air while filling the chamber. The numerical model depicted free surface waves as promoters of air entrainment that ultimately may explain the significant amount of air content in the chamber observed in preliminary tests after the filling process is completed. Results demonstrated that for the present design, against the intuition, the chamber must be filled with liquid at a modest flow rate to minimize free surface waviness during the flooding stage of the chamber

Surface modification of stainless steel for biomedical applications: Revisiting a century-old material

Stainless steel (SS) has been widely used as a material for fabricating cardiovascular stents/valves, orthopedic prosthesis, and other devices and implants used in biomedicine due to its malleability and resistance to corrosion and fatigue. Despite its good mechanical properties, SS (as other metals) lacks biofunctionality. To be successfully used as a biomaterial, SS must be made resistant to the biological environment by increasing its anti-fouling properties, preventing biofilm formation (passive surface modification), and imparting functionality for eluting a specific drug or capturing selected cells (active surface modification); these features depend on the final application. Various physico-chemical techniques, including plasma vapor deposition, electrochemical treatment, and attachment of different linkers that add functional groups, are used to obtain SS with increased corrosion resistance, improved osseointegration capabilities, added hemocompatibility, and enhanced antibacterial properties. Existing literature on this topic is extensive and has not been covered in an integrated way in previous reviews. This review aims to fill this gap, by surveying the literature on SS surface modification methods, as well as modification routes tailored for specific biomedical applications. STATEMENT OF SIGNIFICANCE: Stainless steel (SS) is widely used in many biomedical applications including bone implants and cardiovascular stents due to its good mechanical properties, biocompatibility and low price. Surface modification allows improving its characteristics without compromising its important bulk properties. SS with improved blood compatibility (blood contacting implants), enhanced ability to resist bacterial infection (long-term devices), better integration with a tissue (bone implants) are examples of successful SS surface modifications. Existing literature on this topic is extensive and has not been covered in an integrated way in previous reviews. This review paper aims to fill this gap, by surveying the literature on SS surface modification methods, as well as to provide guidance for selecting appropriate modification routes tailored for specific biomedical applications.Accepted manuscrip

Selection, characterization, and application of DNA aptamers for detection of Mycobacterium tuberculosis secreted protein MPT64

Abstract Rapid detection of Mycobacterium tuberculosis (Mtb), an etiological agent of tuberculosis (TB), is important for global control of this disease. Aptamers have emerged as a potential rival for antibodies in therapeutics, diagnostics and biosensing due to their inherent characteristics. The aim of the current study was to select and characterize single-stranded DNA aptamers against MPT64 protein, one of the predominant secreted proteins of Mtb pathogen. Aptamers specific to MPT64 protein were selected in vitro using systematic evolution of ligands through exponential enrichment (SELEX) method. The selection was started with a pool of ssDNA library with randomized 40-nucleotide region. A total of 10 cycles were performed and seventeen aptamers with unique sequences were identified by sequencing. Dot Blot analysis was performed to monitor the SELEX process and to conduct the preliminary tests on the affinity and specificity of aptamers. Enzyme linked oligonucleotide assay (ELONA) showed that most of the aptamers were specific to the MPT64 protein with a linear correlation of R2 = 0.94 for the most selective. Using Surface Plasmon Resonance (SPR), dissociation equilibrium constant KD of 8.92 nM was obtained. Bioinformatics analysis of the most specific aptamers revealed the existence of a conserved as well as distinct sequences and possible binding site on MPT64. The specificity was determined by testing non-target ESAT-6 and CFP-10. Negligible cross-reactivity confirmed the high specificity of the selected aptamer. The selected aptamer was further tested on clinical sputum samples using ELONA and had sensitivity and specificity of 91.3% and 90%, respectively. Microscopy, culture positivity and nucleotide amplification methods were used as reference standards. The aptamers studied could be further used for the development of medical diagnostic tools and detection assays for Mtb

Multiplexed detection of cancer biomarkers using an optical biosensor

Early detection of cancer is important in administering timely treatment and increasing cancer survival rates. For early cancer detection one can use biomarkers, which are characteristics that can be objectively measured or evaluated as indicators of normal or pathogenic processes. In our study we study three protein biomarkers: carcinoembryonic antigen (CEA), interleukin-6 (IL-6) and extracellular protein kinase A (ECPKA), which have been implicated in various types of human cancer. The main objective of this project is to develop a biosensor for detection of multiple cancer biomarkers. To detect these protein biomarkers high affinity ssDNA aptamers are being selected. Aptamers are short single stranded DNAs with an ability to bind to various targets with high affinity and specificity which selected by SELEX (Systemic Evolution of Ligands through Exponential enrichment) [2]. Ultimately, aptamers against each of the biomarker will be conjugated to magnetic nanoparticles to capture biomarkers from biological fluids. Another aptamer is proposed to be conjugated to quantum dots for quantitation of biomarkers when analyzed on spectrometer

An aptasensor for the detection of Mycobacterium tuberculosis secreted immunogenic protein MPT64 in clinical samples towards tuberculosis detection

This work presents experimental results on detection of Mycobacterium tuberculosis secreted protein MPT64 using an interdigitated electrode (IDE) which acts as a platform for capturing an immunogenic protein and an electrochemical impedance spectroscopy (EIS) as a detection technique. The assay involves a special receptor, single stranded DNA (ssDNA) aptamer, which specifically recognizes MPT64 protein. The ssDNA immobilization on IDE was based on a co-adsorbent immobilization at an optimized ratio of a 1/100 HS-(CH6)(6)-OP(O)(2)O-(CH2CH2O)(6)-5'-TTTTT-aptamer-3'/6-mercaptohexanol. The optimal sample incubation time required for a signal generation on an aptamer modified IDE was found to be at a range of 15-20 min. Atomic Force Microscopy (AFM) results confirmed a possible formation of an aptamer - MPT64 complex with a 20 nm roughness on the IDE surface vs. 4.5 nm roughness for the IDE modified with the aptamer only. A limit of detection for the EIS aptasensor based on an IDE for the detection of MPT64 in measurement buffer was 4.1 fM. The developed EIS aptasensor was evaluated on both serum and sputum clinical samples from the same TB (-) and TB (+) patients having a specificity and sensitivity for the sputum sample analysis 100% and 76.47%, respectively, and for the serum sample analysis 100% and 88.24%, respectively. The developed aptasensor presents a sensitive method for the TB diagnosis with the fast detection time

Optimizing Silanization to Functionalize Stainless Steel Wire: Towards Breast Cancer Stem Cell Isolation

Chemically modified metal surfaces have been used to recognize and capture specific cell types and biomolecules. In this work, stainless steel wires were functionalized with aptamers against breast cancer stem cell markers. Stainless steel wires were first electropolished and silanized via electrodeposition. Aptamers were then attached to the silanized surface through a cross-linker. The functionalized wires were able to capture the target cells in an in vitro test. During surface modification steps, wires were analyzed by atomic force microscopy, cyclic voltammetry, scanning electron and fluorescence microscopy to determine their surface composition and morphology. Optimized conditions of silanization (applied potential, solution pH, heat treatment temperature) for obtaining an aptamer-functionalized wire were determined in this work together with the use of several surface characterization techniques suitable for small-sized and circular wires. These modified wires have potential applications for the in vivo capture of target cells in blood flow, since their small size allows their insertion as standard guidewires in biomedical devices