A descriptive study of lead arm muscle activation patterns during cricket batting
- Authors: McCarthy, Ryan Aidan
- Date: 2024-10-11
- Subjects: Cricket Batting , Muscle contraction , Elbow , Wrist , Forearm , Cricket players , School sports
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/464754 , vital:76542
- Description: Introduction: Kinematic studies identify elbow extension and wrist flexion as key movements for bat swinging speed during cricket batting. Knowledge of lead arm muscle activation during cricket batting may provide a greater understanding of the mechanics leading to the generation of faster bat speed. Aim: The current investigation aimed to determine selected muscle activation of cricketer's lead arm, maximum bat swinging speed and angular separation of the pelvis and thorax in the transverse plane (known as the X-angle) during a distance-hitting batting protocol. Materials and Methods: 12 cricket players playing for a South African school 1st XI school team aged 16-19 years completed a distance-hitting batting protocol. Due to issues in data recording two participants' data was excluded leading to 10 participants being kept in the final analyses.Participants attempted to hit straight lofted drives while facing full-length deliveries at 80-100 km.h-1 from a bowling machine. Each participant faced 12-24 balls split into overs while lead arm muscle activation, bat speed, and angle of lead-elbow flexion were recorded. Each participant's elbow and wrist flexors and extensors were monitored by electromyography (EMG) units while they batted. The EMG units were placed on the wrist flexor and extensor origins and on Triceps and Biceps Brachii. Muscle activation for each muscle was recorded in millivolts (mv) and divided by the maximum measurement of voluntary muscle activation to determine the percentage of maximum voluntary contraction (%MVC) during each shot. Percentage MVC for batting phases of backlift, downswing, contact and follow-through were averaged to compare average %MVC per muscle across all batters and obtain inter-participant variability. Lead arm elbow angle and the %MVC of the elbow and wrist muscles will be compared between successful and unsuccessful shots for analysis. Each participant's lead arm elbow angle was monitored in degrees of flexion by a goniometer placed across the anterior aspect of the elbow joint. Due to the data not having a normal distribution, non-parametric tests were used to establish the variance between dependent variables. To determine the effect of multiple groups on the independent variables a Kruskal-Wallis test for ANOVA was used. Where significant differences were identified, multiple pairwise comparisons were completed to determine where the differences occurred. Results: Successful Lofted straight drives (32 shots) were compared to unsuccessful shots (101 shots) across all participants. Participants and batting phase were found to be significantly different for lead arm elbow angle and muscle activation however shot type was not found to be significantly different. Meaning that lead arm elbow angle and muscle activation differ based on the participant observed or based on the batting phase observed. The backswing phase recorded a higher amount of elbow flexion (155.25°) for Lofted straight drive compared to unsuccessful shots (157.86°) and lower activation across all muscles for Lofted straight drive. The lofted straight drive had a higher amount of elbow flexion (129.52°) compared to unsuccessful shots (149.24°) for the downswing phase and muscle activation was similar with greater variation for unsuccessful shots. At contact Lofted straight drive had a higher amount of elbow flexion (153.44° v 160.13°), and higher activation in the Biceps brachii (34.61% v 28.41%) and Triceps brachii (51.07% v 43.02%). For the follow-through phase Lofted straight drives had a higher amount of elbow flexion (144.87° v 149.59°) and greater Forearm extensor activation (37.13% v 31.28%). There was a large variation across all phases (coefitient of variation between 8.79%-70.28%) with backswing having the least variation and contact having the greatest. Meaning that the backswing phase is fairly predictable for batters and the contact phase is highly variable. Conclusion: Muscle activation increased in the last few milliseconds before contact. The Forearm extensor had the greatest activation during the backswing and follow-through phases. During the downswing phase, Forearm flexors had the greatest activation and at contact, the Triceps brachii had the greatest activation. This study emphasizes the importance of forearm and elbow muscle for batting. Appropriate strengthening of the muscles could also help a batter execute a powerful lofted drive. Future studies with objective measures linked to batting success in prior studies can build on the importance of these findings for batter success. This study provides insight into individual batter techniques and identifies important topics for future research. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2024
- Full Text:
- Authors: McCarthy, Ryan Aidan
- Date: 2024-10-11
- Subjects: Cricket Batting , Muscle contraction , Elbow , Wrist , Forearm , Cricket players , School sports
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/464754 , vital:76542
- Description: Introduction: Kinematic studies identify elbow extension and wrist flexion as key movements for bat swinging speed during cricket batting. Knowledge of lead arm muscle activation during cricket batting may provide a greater understanding of the mechanics leading to the generation of faster bat speed. Aim: The current investigation aimed to determine selected muscle activation of cricketer's lead arm, maximum bat swinging speed and angular separation of the pelvis and thorax in the transverse plane (known as the X-angle) during a distance-hitting batting protocol. Materials and Methods: 12 cricket players playing for a South African school 1st XI school team aged 16-19 years completed a distance-hitting batting protocol. Due to issues in data recording two participants' data was excluded leading to 10 participants being kept in the final analyses.Participants attempted to hit straight lofted drives while facing full-length deliveries at 80-100 km.h-1 from a bowling machine. Each participant faced 12-24 balls split into overs while lead arm muscle activation, bat speed, and angle of lead-elbow flexion were recorded. Each participant's elbow and wrist flexors and extensors were monitored by electromyography (EMG) units while they batted. The EMG units were placed on the wrist flexor and extensor origins and on Triceps and Biceps Brachii. Muscle activation for each muscle was recorded in millivolts (mv) and divided by the maximum measurement of voluntary muscle activation to determine the percentage of maximum voluntary contraction (%MVC) during each shot. Percentage MVC for batting phases of backlift, downswing, contact and follow-through were averaged to compare average %MVC per muscle across all batters and obtain inter-participant variability. Lead arm elbow angle and the %MVC of the elbow and wrist muscles will be compared between successful and unsuccessful shots for analysis. Each participant's lead arm elbow angle was monitored in degrees of flexion by a goniometer placed across the anterior aspect of the elbow joint. Due to the data not having a normal distribution, non-parametric tests were used to establish the variance between dependent variables. To determine the effect of multiple groups on the independent variables a Kruskal-Wallis test for ANOVA was used. Where significant differences were identified, multiple pairwise comparisons were completed to determine where the differences occurred. Results: Successful Lofted straight drives (32 shots) were compared to unsuccessful shots (101 shots) across all participants. Participants and batting phase were found to be significantly different for lead arm elbow angle and muscle activation however shot type was not found to be significantly different. Meaning that lead arm elbow angle and muscle activation differ based on the participant observed or based on the batting phase observed. The backswing phase recorded a higher amount of elbow flexion (155.25°) for Lofted straight drive compared to unsuccessful shots (157.86°) and lower activation across all muscles for Lofted straight drive. The lofted straight drive had a higher amount of elbow flexion (129.52°) compared to unsuccessful shots (149.24°) for the downswing phase and muscle activation was similar with greater variation for unsuccessful shots. At contact Lofted straight drive had a higher amount of elbow flexion (153.44° v 160.13°), and higher activation in the Biceps brachii (34.61% v 28.41%) and Triceps brachii (51.07% v 43.02%). For the follow-through phase Lofted straight drives had a higher amount of elbow flexion (144.87° v 149.59°) and greater Forearm extensor activation (37.13% v 31.28%). There was a large variation across all phases (coefitient of variation between 8.79%-70.28%) with backswing having the least variation and contact having the greatest. Meaning that the backswing phase is fairly predictable for batters and the contact phase is highly variable. Conclusion: Muscle activation increased in the last few milliseconds before contact. The Forearm extensor had the greatest activation during the backswing and follow-through phases. During the downswing phase, Forearm flexors had the greatest activation and at contact, the Triceps brachii had the greatest activation. This study emphasizes the importance of forearm and elbow muscle for batting. Appropriate strengthening of the muscles could also help a batter execute a powerful lofted drive. Future studies with objective measures linked to batting success in prior studies can build on the importance of these findings for batter success. This study provides insight into individual batter techniques and identifies important topics for future research. , Thesis (MSc) -- Faculty of Science, Human Kinetics and Ergonomics, 2024
- Full Text:
Teacher professional development as a mechanism to advance education for sustainable development (ESD) integration in Namibia: a senior primary English language case study
- Shangheta, Miryam Keshityeni
- Authors: Shangheta, Miryam Keshityeni
- Date: 2024-10-11
- Subjects: Career development , Teachers In-service training , Education for sustainable development , English language Study and teaching Namibia , Pedagogical content knowledge
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/463603 , vital:76424
- Description: Namibia is a signatory to the Sustainable Development Goals of Agenda 2030 and responds to the United Nations Decade of Education for Sustainable Development by incorporating Education for Sustainable Development (ESD) into its education curriculum and policies. In 2020, Namibia launched its National Environmental Education and Education for Sustainable Development Policy. Despite these policy commitments, research indicates that most teachers continue to struggle with integrating ESD into their teaching practices. The integration of ESD in English language teaching classrooms is no exception. This qualitative case study is an attempt to understand how a Teacher Professional Development programme named Teach for ESD offered by NaDEET, a non-governmental organisation in Namibia, aided three Senior Primary Phase English Language teachers to integrate ESD into their teaching practice. The study also aims to unveil the teaching practice of the selected English Language teachers in the Otjozondjupa region who participated in the Teach for ESD programme. Data was gathered using semi-structured interviews (three interviews per teacher), document review, and lesson observation (one per teacher). Theoretically, the study is framed by socio-cultural learning theory after the work of Lev Vygotsky, supplemented by Mavhunga and Rollnick’s (2013) account of Topic Specific Pedagogical Content Knowledge (TSPCK). The qualitative data was analysed inductively to identify themes and sub-themes relevant to the research questions. The study findings might be of value to NaDEET as they implement future TPD courses in ESD, and they may also be of interest to English Language Teaching stakeholders as Namibia implements the EE and ESD Policy in the national school curriculum. The findings showed that the English Language teachers need ESD training; ESD-oriented teaching and learning materials; continuing support for monitoring and evaluation of their ESD practices; and more time allocated to English Language teaching in the timetable so that the environmental / sustainability content can be explored in more depth alongside the development of English Language skills. The study recommends that further research be conducted into effective ways of teaching English Language skills through environmental themes (developing Pedagogical Content Knowledge); and exploring up-scalable models of teacher professional development so that more English Language teachers can receive ESD training. , Thesis (MEd) -- Faculty of Education, Secondary and Post-School Education, 2024
- Full Text:
- Authors: Shangheta, Miryam Keshityeni
- Date: 2024-10-11
- Subjects: Career development , Teachers In-service training , Education for sustainable development , English language Study and teaching Namibia , Pedagogical content knowledge
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/463603 , vital:76424
- Description: Namibia is a signatory to the Sustainable Development Goals of Agenda 2030 and responds to the United Nations Decade of Education for Sustainable Development by incorporating Education for Sustainable Development (ESD) into its education curriculum and policies. In 2020, Namibia launched its National Environmental Education and Education for Sustainable Development Policy. Despite these policy commitments, research indicates that most teachers continue to struggle with integrating ESD into their teaching practices. The integration of ESD in English language teaching classrooms is no exception. This qualitative case study is an attempt to understand how a Teacher Professional Development programme named Teach for ESD offered by NaDEET, a non-governmental organisation in Namibia, aided three Senior Primary Phase English Language teachers to integrate ESD into their teaching practice. The study also aims to unveil the teaching practice of the selected English Language teachers in the Otjozondjupa region who participated in the Teach for ESD programme. Data was gathered using semi-structured interviews (three interviews per teacher), document review, and lesson observation (one per teacher). Theoretically, the study is framed by socio-cultural learning theory after the work of Lev Vygotsky, supplemented by Mavhunga and Rollnick’s (2013) account of Topic Specific Pedagogical Content Knowledge (TSPCK). The qualitative data was analysed inductively to identify themes and sub-themes relevant to the research questions. The study findings might be of value to NaDEET as they implement future TPD courses in ESD, and they may also be of interest to English Language Teaching stakeholders as Namibia implements the EE and ESD Policy in the national school curriculum. The findings showed that the English Language teachers need ESD training; ESD-oriented teaching and learning materials; continuing support for monitoring and evaluation of their ESD practices; and more time allocated to English Language teaching in the timetable so that the environmental / sustainability content can be explored in more depth alongside the development of English Language skills. The study recommends that further research be conducted into effective ways of teaching English Language skills through environmental themes (developing Pedagogical Content Knowledge); and exploring up-scalable models of teacher professional development so that more English Language teachers can receive ESD training. , Thesis (MEd) -- Faculty of Education, Secondary and Post-School Education, 2024
- Full Text:
Formulation of an enzyme cocktail, HoloMix, using cellulolytic and xylanolytic enzyme core-sets for effective degradation of various pre-treated hardwoods
- Authors: Malgas, Samkelo
- Date: 2018
- Subjects: Biomass , Cellulase , Hardwoods , Xylanases
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/62827 , vital:28297 , DOI https://doi.org/10.21504/10962/62827
- Description: Currently, there is a growing interest in utilising hardwoods as feedstocks for bioethanol production due to the vast advantages they have over other feedstocks for fermentable sugar production. In this study, two selected hardwoods, Acacia and Populus spp., were subjected to two pre-treatment processes (Sodium chlorite delignification and Steam explosion) and compared with respect to how these pre-treatments affect their enzymatic saccharification. Hardwoods were selected for this study, because hardwoods are easier to delignify when compared to softwoods, and therefore their polysaccharides are more easily accessible by enzymes for the purpose of producing fermentable sugars. Currently available commercial enzyme mixtures have been developed for optimal hydrolysis of acid-pre-treated corn stover and are therefore not optimal for saccharification of pre-treated hardwoods. In this work, we attempted the empirical design of a hardwood specific enzyme cocktail, HoloMix. Firstly, a cellulolytic core-set, CelMix (in a ratio of Egl 68%: Cel7A 17%: Cel6A 6%: Bgl1 9%), for the optimal release of glucose, and a xylanolytic core-set, XynMix (in a ratio of Xyn2A 60%: XT6 20%: AguA 11%: SXA 9%), for the optimal release of xylose, were formulated using an empirical enzyme ratio approach after biochemically characterising these enzymes. As it is well ̶ known that biomass pre-treatment may result in the generation of compounds that hamper enzymatic hydrolysis and microbial fermentation, the effects of these compounds on CelMix and XynMix were evaluated. Using the optimised CelMix and XynMix cocktails, a HoloMix cocktail was established for optimal reducing sugar, glucose and xylose release from the various pre-treated hardwoods. For delignified biomass, the optimized HoloMix consisted of CelMix to XynMix at 75% to 25% protein loading, while for the untreated and steam exploded biomass the HoloMix consisted of CelMix to XynMix at 93.75% to 6.25% protein loading. Sugar release by the HoloMix at a loading of 27.5 mg protein/g of biomass (or 55 mg protein/g of glucan) after 24 h gave 70-100% sugar yield. Treatment of the hardwoods with a laccase from Agaricus bisporus, especially wood biomass with a higher proportion of lignin, significantly improved saccharification by the formulated HoloMix enzyme cocktails. This study provided insights into the enzymatic hydrolysis of various pre-treated hardwood substrates and assessed whether the same lignocellulolytic cocktail can be used to efficiently hydrolyse different hardwood species. The present study also demonstrated that the hydrolysis efficiency of the optimised HoloMix was comparable to (if not better) than commercial enzyme preparations during hardwood biomass saccharification. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Authors: Malgas, Samkelo
- Date: 2018
- Subjects: Biomass , Cellulase , Hardwoods , Xylanases
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/62827 , vital:28297 , DOI https://doi.org/10.21504/10962/62827
- Description: Currently, there is a growing interest in utilising hardwoods as feedstocks for bioethanol production due to the vast advantages they have over other feedstocks for fermentable sugar production. In this study, two selected hardwoods, Acacia and Populus spp., were subjected to two pre-treatment processes (Sodium chlorite delignification and Steam explosion) and compared with respect to how these pre-treatments affect their enzymatic saccharification. Hardwoods were selected for this study, because hardwoods are easier to delignify when compared to softwoods, and therefore their polysaccharides are more easily accessible by enzymes for the purpose of producing fermentable sugars. Currently available commercial enzyme mixtures have been developed for optimal hydrolysis of acid-pre-treated corn stover and are therefore not optimal for saccharification of pre-treated hardwoods. In this work, we attempted the empirical design of a hardwood specific enzyme cocktail, HoloMix. Firstly, a cellulolytic core-set, CelMix (in a ratio of Egl 68%: Cel7A 17%: Cel6A 6%: Bgl1 9%), for the optimal release of glucose, and a xylanolytic core-set, XynMix (in a ratio of Xyn2A 60%: XT6 20%: AguA 11%: SXA 9%), for the optimal release of xylose, were formulated using an empirical enzyme ratio approach after biochemically characterising these enzymes. As it is well ̶ known that biomass pre-treatment may result in the generation of compounds that hamper enzymatic hydrolysis and microbial fermentation, the effects of these compounds on CelMix and XynMix were evaluated. Using the optimised CelMix and XynMix cocktails, a HoloMix cocktail was established for optimal reducing sugar, glucose and xylose release from the various pre-treated hardwoods. For delignified biomass, the optimized HoloMix consisted of CelMix to XynMix at 75% to 25% protein loading, while for the untreated and steam exploded biomass the HoloMix consisted of CelMix to XynMix at 93.75% to 6.25% protein loading. Sugar release by the HoloMix at a loading of 27.5 mg protein/g of biomass (or 55 mg protein/g of glucan) after 24 h gave 70-100% sugar yield. Treatment of the hardwoods with a laccase from Agaricus bisporus, especially wood biomass with a higher proportion of lignin, significantly improved saccharification by the formulated HoloMix enzyme cocktails. This study provided insights into the enzymatic hydrolysis of various pre-treated hardwood substrates and assessed whether the same lignocellulolytic cocktail can be used to efficiently hydrolyse different hardwood species. The present study also demonstrated that the hydrolysis efficiency of the optimised HoloMix was comparable to (if not better) than commercial enzyme preparations during hardwood biomass saccharification. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
The creation and validation of aptamers binding to murine 3T3-L1 Preadipocytes: preliminary implications for controlled cellular attachment, differentiation and cell fate
- Authors: Rubidge, Mark Lourens
- Date: 2017
- Subjects: Oligonucleotides , Fat cells , Stem cells , Ligand binding (Biochemistry) , Fluorimetry
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65247 , vital:28714
- Description: The controlled seeding of a variety of stem cells in vitro has been reported to alter the patterns of their subsequent differentiation. This has been attributed to the control of the surface microenvironment onto which adherent stem cells are cultured, especially control of the proximal density of neighbouring cells. Simultaneously, advances in the generation of aptamers - synthetic ligand molecules developed using in vitro selection techniques targeting complex molecules - have aided in the production of molecules capable of selectively binding to a variety of commercial stem cell lines. Combining the aforementioned research fields, the project reported in this thesis aimed to generate DNA-based aptamers capable of assisting with the selective binding of murine 3T3- L1 preadipocytes to a solid surface. This was performed with a view to, eventually, control the seeding densities of the adherent preadipocytes on the surface of the tissue culture dish in subsequent researchers. In the process of meeting this goal, several optimisations of the in vitro process by which aptamers binding to cells are generated (Cell-SELEX) were performed: an analysis into a variety of methods used for the removal of the single stranded aptamer candidate sequences attached to the surface of 3T3-L1 preadipocytes, a comparison of methods for the generation of single-stranded aptamer sequences from double-stranded DNA template molecules and a method for quantifying the removed ssDNA from the cell surface. Their use is further reported in this work. Initially, it was determined that a fluorimetric evaluation of the unbound single stranded DNA was the optimum technique to use to evaluate the relative amounts of aptamer DNA binding to target cells during cell-SELEX; this arose from the release of DNA, and other cell lysate contaminates, which interfered UV/ Vis quantification. The evaluation into different methods of ssDNA removal from the cell surface showed that although trypsinisation of the cells demonstrated the highest level of aptamer detachment (quantified by fluorimetry), there is a decrease the number of potential targets that aptamers could attach to. The most common method for detaching bound DNA aptamer molecules from cellular targets reported in literature, the use of high temperatures, was selected for cell-SELEX to increase the variability in potential target sites on the cell surface. Using techniques optimised in this work, fluorescently-tagged single-stranded oligonucleotide aptamers were later generated with a positive selection pressure to bind to the surface of the 3T3-L1 preadipocytes, but not to their differentiated adipocyte counterparts. After eight cycles of cell-SELEX, fluorescent spectroscopic analysis depicted a 74 % binding retention of the selection pool in the positive preadipocyte selection pool, as opposed to a 0.69 % binding of sequences to the negative differentiated preadipocytes. Following the isolation and identification of candidate sequences, seven separate sequences were identified as being successfully generated from the selection process. Bioinformatic characterisation of these placed sequenced aptamer candidates into two separate families, that were then analysed in opposition to each for their binding affinity toward each other. Using fluorescently-tagged sequences, the binding selectivity of the generated aptamers was validated using both epifluorescent microscopy and confocal microscopy. At this stage, an aptamer sequence selected from prior in-house research to serve as a negative control also demonstrated significant binding to the extracellular matrix of both preadipocytes and mature adipocytes. 5’-thiolated aptamer sequences were used to form self-assembled monolayers on the electrode surfaces of the impedimetric Roche xCELLigence Real-Time Cell Analysis. The use of aptamer sequences to capture the seeded preadipocytes, demonstrated a slight increase in the extent of binding of the preadipocytes to the gold electrode surface and produced some preliminary indications of alterations to the pattern and rate of subsequent differentiation in the preadipocytes. This provides preliminary evidence that aptamers developed to bind specifically to a stem cell line in vitro show potential to be used as to capture said cell when cast in a self- assembled monolayer assembly. This provides a future opportunity to control the seeding densities of the cells in vitro. The effects of cellular differentiation at a set of predefined cellular densities can be demonstrated on a desired stem cell line. , Thesis (MSc) -- Faculty of Faculty of Science, Biotechnology Innovation Centre, 2017
- Full Text:
- Authors: Rubidge, Mark Lourens
- Date: 2017
- Subjects: Oligonucleotides , Fat cells , Stem cells , Ligand binding (Biochemistry) , Fluorimetry
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/65247 , vital:28714
- Description: The controlled seeding of a variety of stem cells in vitro has been reported to alter the patterns of their subsequent differentiation. This has been attributed to the control of the surface microenvironment onto which adherent stem cells are cultured, especially control of the proximal density of neighbouring cells. Simultaneously, advances in the generation of aptamers - synthetic ligand molecules developed using in vitro selection techniques targeting complex molecules - have aided in the production of molecules capable of selectively binding to a variety of commercial stem cell lines. Combining the aforementioned research fields, the project reported in this thesis aimed to generate DNA-based aptamers capable of assisting with the selective binding of murine 3T3- L1 preadipocytes to a solid surface. This was performed with a view to, eventually, control the seeding densities of the adherent preadipocytes on the surface of the tissue culture dish in subsequent researchers. In the process of meeting this goal, several optimisations of the in vitro process by which aptamers binding to cells are generated (Cell-SELEX) were performed: an analysis into a variety of methods used for the removal of the single stranded aptamer candidate sequences attached to the surface of 3T3-L1 preadipocytes, a comparison of methods for the generation of single-stranded aptamer sequences from double-stranded DNA template molecules and a method for quantifying the removed ssDNA from the cell surface. Their use is further reported in this work. Initially, it was determined that a fluorimetric evaluation of the unbound single stranded DNA was the optimum technique to use to evaluate the relative amounts of aptamer DNA binding to target cells during cell-SELEX; this arose from the release of DNA, and other cell lysate contaminates, which interfered UV/ Vis quantification. The evaluation into different methods of ssDNA removal from the cell surface showed that although trypsinisation of the cells demonstrated the highest level of aptamer detachment (quantified by fluorimetry), there is a decrease the number of potential targets that aptamers could attach to. The most common method for detaching bound DNA aptamer molecules from cellular targets reported in literature, the use of high temperatures, was selected for cell-SELEX to increase the variability in potential target sites on the cell surface. Using techniques optimised in this work, fluorescently-tagged single-stranded oligonucleotide aptamers were later generated with a positive selection pressure to bind to the surface of the 3T3-L1 preadipocytes, but not to their differentiated adipocyte counterparts. After eight cycles of cell-SELEX, fluorescent spectroscopic analysis depicted a 74 % binding retention of the selection pool in the positive preadipocyte selection pool, as opposed to a 0.69 % binding of sequences to the negative differentiated preadipocytes. Following the isolation and identification of candidate sequences, seven separate sequences were identified as being successfully generated from the selection process. Bioinformatic characterisation of these placed sequenced aptamer candidates into two separate families, that were then analysed in opposition to each for their binding affinity toward each other. Using fluorescently-tagged sequences, the binding selectivity of the generated aptamers was validated using both epifluorescent microscopy and confocal microscopy. At this stage, an aptamer sequence selected from prior in-house research to serve as a negative control also demonstrated significant binding to the extracellular matrix of both preadipocytes and mature adipocytes. 5’-thiolated aptamer sequences were used to form self-assembled monolayers on the electrode surfaces of the impedimetric Roche xCELLigence Real-Time Cell Analysis. The use of aptamer sequences to capture the seeded preadipocytes, demonstrated a slight increase in the extent of binding of the preadipocytes to the gold electrode surface and produced some preliminary indications of alterations to the pattern and rate of subsequent differentiation in the preadipocytes. This provides preliminary evidence that aptamers developed to bind specifically to a stem cell line in vitro show potential to be used as to capture said cell when cast in a self- assembled monolayer assembly. This provides a future opportunity to control the seeding densities of the cells in vitro. The effects of cellular differentiation at a set of predefined cellular densities can be demonstrated on a desired stem cell line. , Thesis (MSc) -- Faculty of Faculty of Science, Biotechnology Innovation Centre, 2017
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