Doctor of Philosophy, University of Akron, 2024, Polymer Engineering

In this work, High Density Polyethylene (HDPE) composites were made using Torrefied Soybean Hulls (TSBH) and Carbon Black (CB) to study the interactions affiliated with the TSBH content for as-received as well as size-reduced particles. The Milled TSBH (MTSBH) was shown to integrate well at low loadings, but showed signs of favoring filler-filler interactions over filler-matrix interactions, reducing the overall effectiveness as the loadings increased. Rheological testing showed that the higher-loaded MTSBH composites behaved similar to composites with larger particles as the loading increased, indicating that clusters had formed. Unmilled TSBH (UTSBH) showed good mechanical strength, but the particle size was shown to limit its ability to integrate into the material, even at low loadings. The addition of CB was shown to have the most impact on the low loading MTSBH composites, where the MTSBH-CB interactions were shown to influence the filler network in electrical resistance testing where a nonlinear trend was observed in the composite resistivity with the addition of MTSBH. In UTSBH composites, there were less signs of CB-UTSBH interactions due to the relatively large particle size. To contrast the hydrophilic matrix behavior of HDPE, Nylon-6 (PA6) was used as a matrix for the TSBH composites. In cases where either TSBH filler was used, the composite performance was shown to improve to a greater degree than in the case of HDPE due to the hydrophilic groups contained in the PA6 backbone. Similar to the HDPE composites, the TSBH particles showed a lack of effectiveness at higher filler loadings, though MTSBH showed more effective integration which indicates that this is a result of particle size. The CB and MTSBH showed synergistic effects with high CB and low MTSBH loading during cyclic tension testing, where the increase in strain energy density required for a test was less when the CB was present that when it was not. This effect was seen throughout the mono (open full item for complete abstract)

Doctor of Philosophy (PhD), Ohio University, 2022, Chemistry and Biochemistry (Arts and Sciences)

Non-covalent interactions govern biological processes, self-assembling systems, complex materials, and molecular machinery. Cucurbit[n]urils (CB[n]s) are pumpkin-shaped, water-soluble molecular containers that have a hydrophobic cavity and two hydrophilic carbonylated portals. This research focuses on the design and synthesis of various CB[8]-secured supramolecular assemblies to study and quantify non-covalent interactions in the aqueous medium. We first report the design of a CB[8]-based hydrocarbon sensitive chemosensor, where the selectivity is exclusively driven by dispersive interactions between the hydrocarbons and the cavity formed by a portion of the inner wall of CB[8] and the positive auxiliary guest. We also proposed an empirical model which can predict the binding selectivity of hydrocarbons and noble gases to CB[n] analogs when the macrocycle is mimicked with a non-polar organic solvent. We then synthesized various CB[8]-mediated supramolecular architectures as possible brakes and gears driven by favorable π-π and metal-metal interactions. Finally, we proposed a supramolecular tool (a molecular balance) to quantify non-covalent interactions in the aqueous medium.

Doctor of Philosophy, The Ohio State University, 2008, Biophysics

The most controversial step in the study of the mechanism of action of insecticidal crystal toxins is that of insertion of the toxin into insect brush border membranes. Conflicting models of insertion of toxin can be categorized into two groups; ones that propose that only certain alpha helices of domain I insert into insect brush border membrane vesicles (BBMV) (Umbrella, Penknife and Serial Receptor Binding models) and others that propose that most of the toxin inserts into BBMV (Aronson, Buried Dragon and Unchanged Structure models). Protease protection studies of cysteine mutations from all domains of the toxin showed protection of most of the toxin (a 60 kDa form) similar to the wild type Cry1A toxin, when inserted into insect brush border membranes. Studies on steady state fluorescence measurements of these cysteine residues when bound to artificial vesicles or natural brush border membrane vesicles (BBMV), and fluorescence energy transfer measurements in labeled artificial vesicles suggested that residues from all the domains of the toxin inserted into the membrane. Residues in the loop 2 of Domain II of the toxin that played a vital role in the insertion of the toxin into insect BBMV were identified. Examination of receptor binding and insertion of mutants of these residues have shown that insertion of Cry1Ab into the membrane is dependent on specific residues at positions in this loop. Absence of phenylalanine or a closely related amino acid such as tryptophan at position 371 allowed initial binding of the toxin to the receptor but compromised the insertion of the toxin into insect membrane, thereby confirming that irreversible binding step of the toxin-BBMV interaction is the critical step in the mode of action of the toxin where Domain II is a major candidate mediating the step. Fluorescence blue shift studies into artificial and natural membranes also indicated a difference in the partitioning of the toxin into artificial and natural membranes; thereby s (open full item for complete abstract)

Master of Sciences (Engineering), Case Western Reserve University, 2024, EECS - Computer and Information Sciences

The clustering of single-cell RNA-sequencing data has been established as an important first step in single-cell gene expression data analysis for scientists to identify cell type based on RNA level expression. This is important because once a cell type has been identified, the phenotype association, as well as the spatiotemporal dynamics of specific cell types, can be characterized, which could lead to identifying cells associated with cancers and other diseases. However, the high-dimensionality of the data poses computational challenges, while drop-outs (genes that are not identified despite being expressed) hamper the reliability of inference. Since established knowledge on transcriptional regulatory networks provide information on the regulatory relationships between genes, we hypothesize that regulatory networks can help remedy missing data, while also reducing dimensionality. To test this hypothesis, we use a previously existing regulatory network, modern clustering methods, and network propagation together to help enhance clustering performance, which enhances accurate identification of cell types.

Master of Science (MS), Wright State University, 2023, Biological Sciences

Invasive species threaten ecosystems and economies. Globally, biological invasions are estimated to have cost over $2.1 trillion since 1970. In Eastern North American woodlands, invasive plants are rapidly displacing natives. This is concerning, because invasive plants may not support the diverse and abundant arthropod communities essential to ecosystem function. Despite the conceptual understanding of invasive shrubs' potential to transform forest communities, scant research has focused on the effect of invasive plants on higher trophic levels here in Ohio. To address this gap, I examined the diversity and abundance of arthropod communities, caterpillar performance, and caterpillar predation on two invasive shrubs, Amur honeysuckle (Lonicera maackii) and border privet (Ligustrum obtusifolium), relative to native counterparts. Lo. maackii supported a depauperate community; however, Li. obtusifolium hosted a surprisingly robust community. Nevertheless, both invasives proved poor hosts for caterpillars and were less preferred by foraging insectivores.

Master of Science, The Ohio State University, 2023, Animal Sciences

Animals are often used as educational resources in post-secondary educational settings across the United States, especially in animal science curricula. Yet, little is known about the effect of student-animal interactions (SAI) on students' attitudes, curiosity, and empathy (ACE) towards animals or how their demographic backgrounds, past animal experience, and previous animal science coursework affect these areas. Moreover, while these SAI impact the animals used as educational resources, there is minimal understanding regarding their effect on the animals' welfare. Thus, this study aimed to (1) determine if animal sciences students' ACE responses toward animals changed over the course of the semester while identifying any demographic factors contributing to this change and (2) evaluate the emotional states of the animals used as educational resources through measurements of behavioral responses observed during SAI. This study was conducted at The Ohio State University during Autumn semester of 2022. A pre-course (n = 215) and post-course (n = 96) survey was administered to animal sciences students to collect self-reported demographic information, prior animal experience, prior animal science coursework, and ACE responses toward animals by species to determine if there was a change over the semester. Curiosity increased between the pre-course and post-course survey (p < 0.001), with students' attitude, experience, and perceived knowledge of animals (p < 0.001) providing plausible explanations. There was a strong positive correlation between the students' perceived knowledge of species and experience with species, indicating potential for further studies to assess changes in knowledge before and after working with animals (R = 0.793, p < 0.001). The emotional state of animals used as educational resources varied depending on the level of invasiveness of the activity as well as the length of SAI. Animals exhibited a higher frequency of negative emotional states during (open full item for complete abstract)

Master of Science, The Ohio State University, 2023, Evolution, Ecology and Organismal Biology

Restoration is a global priority that aims to reinstate ecosystem function. One key metric of ecosystem function is species interactions. However, the species interactions that promote function can be influenced by land-use legacy, the lasting impact of land-use on the ecosystem. Surface mining is an example of an intensive land-use practice that dramatically alters the abiotic environment, potentially causing an array of legacy effects. Reclamation standards, such as those specified in the Surface Mining Control and Reclamation Act of 1977 (SMCRA), have been introduced over time to mitigate the damage of surface mining. Areas mined prior to SMCRA were reclaimed with few requirements and little consistency, while SMCRA mandated topsoil replacement, contouring, and revegetation. It is unknown if different reclamation techniques have different legacy effects. The overarching goal of this study was to determine if species interactions differ between surface mine reclamation histories. First, to characterize the abiotic differences between reclamation histories, I measured the soil quality of five sites mined pre-SMCRA and five sites mined post-SMCRA in Eastern Ohio. To investigate how reclamation history affects a mutualistic belowground interaction, I planted legumes for eight weeks and observed plant growth and colonization by rhizobia. I also studied an antagonistic aboveground interaction, insect herbivory, by sampling these same plants for foliar damage to determine herbivore preference for plants grown in either reclamation history. I found that pre-SMCRA soils had significantly higher nitrogen, and that nitrogen availability drove plant survival and plant growth. Despite pre-SMCRA sites having higher soil nitrogen content, and previous studies showing stronger associations with microbes in nutrient poor-soils, pre-SMCRA plants had greater rhizobia colonization, suggesting that the increased mutualism in pre-SMCRA sites may be driven by another environmental f (open full item for complete abstract)

Doctor of Philosophy, University of Akron, 2021, Polymer Science

Protein therapeutics are involved in several diseases, including HIV, cancer, and infectious diseases. However, the major challenge is preserving the activity of the protein outside its native environment. The effectiveness of protein therapeutics would be improved by developing a polymeric shell-like structure to encapsulate and release them sustainably. Our lab has developed a new class of non-ionic, thermoresponsive polyesters (TR-PEs), which can create protein delivery cargos. TR-PEs can undergo reversible temperature-dependent phase separation above lower critical solution temperature (LCST) to form coacervates. To test the ability of this coacervate platform, the encapsulation and release of Doxorubicin (Dox), an anti-cancer drug, were analyzed to understand molecular-level interactions with our TR-PEs. Several spectroscopy techniques, including 2D NMR, were employed to probe the coacervate-Dox interactions, and we have shown specific polymer-Dox interactions that lead to enhanced encapsulation. Although the above approach is promising, efficient protein encapsulation requires tailored protein-TR-PE interactions for each protein-TR-PE pair. However, synthesizing a large number of TR-PEs and examining their interactions with protein can be time-consuming. Therefore, a systematic, ‘fragment-based' approach was applied, similar to the pharmaceutical industry's fragment-based drug discovery process. The monomer ‘fragments' of the TR-PEs, methyl esters of the pendant functionalized diols, were developed for this methodology. A blueprint was established for ideal TR-PE compositions with significant binding affinities towards the protein, ubiquitin, by an iterative fragment identification and optimization of protein-fragment interactions. Advanced NMR techniques were exploited in fragment-based TR-PE discovery strategy. This approach can be transformational in designing polymers with specificity towards protein targets.

Doctor of Business Administration (D.B.A.), Franklin University, 2021, Business Administration

Transformational leadership is identified as an innovative managerial approach that highlights empowerment, inspiration, and stimulation of followers (Wang, Kim, & Lee, 2016; Belschak, Den Hartog, & Kalshoven, 2015). Literature supports the idea that a transformational approach to leadership drives innovation and change and creates an environment where employees are inspired and encouraged (Lukowki, 2017). However, the focus of the literature and current research surrounding transformational leadership emphasizes the practical applications utilized within a corporate context, while research analyzing the academic forms of transformational leadership is minimal in comparison. The purpose of this study is to explore a possible relationship between the existence of behaviors by university instructors aligned with transformational leadership and whether these behaviors promote specific student results. The objective of this study is to quantify a correlational relationship between leadership behaviors exhibited by instructors and positive student outcomes, specifically the integration of an interactive teaching style, challenging of students, encouragement of independent thought, and the overall student intellectual stimulation score. The researcher used two existing survey instruments, the Multifactor Leadership Questionnaire (MLQ) (Bass & Avolio, 1995) and the Student Intellectual Stimulation Scale (SISS) (Bolkan & Goodboy, 2010), to test the presence of this correlation.

Master of Sciences, Case Western Reserve University, 2018, Physiology and Biophysics

Erythropoietin-producing human hepatocellular (Eph) receptors account for the majority of the membrane-bound receptor tyrosine kinase (RTK) family. Eph receptors have significant roles during embryonic development, cell maturation, and adulthood. Furthermore, studies have shown that drastic increase in the expression of Eph receptors is detected in the malignant proliferation of tumor cells in numerous cancers, such as melanoma, breast cancer and glioblastoma. Indeed, recently, oncogenic activities have been observed in non-canonical unliganded Ephrin type-A receptor 2 (EphA2). This project aims to probe the effects of phosphorylation on the intracellular domain interactions of EphA2 in solution. Results from this study indicate that deletion of the sterile a motif (SAM) domain leads to a strong binding affinity between kinase domains in solution. Upon oligomerization, less kinase activity is observed, compared to that of the monomeric state of the intracellular domain (ICD) of EphA2. Moreover, in this study, microscale thermophoresis (MST) is utilized to assess the binding interactions of EphA2 in complex with liposomes, a membrane-like environment. Upon the truncation of the SAM domain, binding of the intracellular domains of EphA2 to the membrane is abrogated. Data from this study will help to elucidate and further our understanding of the signaling mechanism of receptor tyrosine kinases and the regulatory role of phosphorylation.

Doctor of Philosophy, The Ohio State University, 1974, Graduate School

Master of Arts (MA), Bowling Green State University, 2016, Sociology

In the United States the share of middle-age and older adults who are stepparents has increased over the recent decades. Prior studies have shown that stepparents generally experience more depressive symptoms than parents with only biological children, but most of this literature has focused on stepparents of minor children. The few studies that have examined the relation between stepparenthood and depressive symptoms in later life find either a positive or a null association. The mixed findings could be attributable to different definitions of stepparenthood used in these studies. Individuals can become simple stepparents when they have only stepchildren or they can become complex stepparents when they have both biological children and stepchildren. To fill this gap in the literature, I used the 2008 and 2010 waves of Health and Retirement Study to examine how simple stepparents, complex stepparents, and parents with only biological children differ in their depressive symptoms, with particular attention given to the mediating roles of parent-child contact and interactions. Women and men were also analyzed separately in this study. The results showed that stepparenthood was not associated with depressive symptoms for women and men. Contact and interactions with children did not mediate the association between parental status and depressive symptoms for either men or women. Potential avenues for future stepfamily research is discussed.

PhD, University of Cincinnati, 2016, Allied Health Sciences: Communication Sciences and Disorders

The purpose of this dissertation research was to describe children's pragmatic skills after traumatic brain injury (TBI) within the context of parent-child interactions. Children with TBI are known to exhibit a variety of pragmatic deficits. How these deficits affect a child's ability to interact with parents is largely unknown. Research examining parent and child pragmatic behavior after TBI is needed because parent-child interactions are an important component of a child's recovery. Two exploratory studies were conducted. Participants included ten mothers and their child with TBI. Children were 6-12 years old, had sustained a moderate to severe TBI, and were more than one-year post-injury. All participants participated in both studies. The first study used a phenomenological qualitative approach to describe mothers' experiences communicating with their child with TBI. The mothers' experiences were collected through semi-structured interviews and questionnaires. Interviews were analyzed using a deductive framework and constant comparison analysis to develop home and outside of the home social contexts and pragmatic deficit themes. The results of the first study showed that mothers primarily described their child with TBI as exhibiting average or near average pragmatic skills at home. However, the majority of mothers observed some problematic communication behaviors resulting in six home and five outside of the home contexts in which pragmatic deficits were observed. These contexts consisted of different pragmatic deficit themes, with only three of ten themes overlapping across environments (home and outside of the home). The second study used a descriptive research design to characterize parent-child pragmatic behaviors during a conversation in the home. During the home visit, children completed language, pragmatics, and general intellect assessments. Mothers completed questionnaires on family functioning and the child's executive function behaviors. Mo (open full item for complete abstract)

Master of Sciences, Case Western Reserve University, 0, Materials Science and Engineering

Proteins and viral nanoparticles are important building blocks for the construction of micro- and nano-scale materials. The behavior of these biomolecular systems in crystallization, self-association and molecular recognition, is determined by the long-range interactions between them. Understanding the variation of long-range interactions under different solution conditions facilitates the manipulation and design of novel hierarchical structures. The second virial coefficient is the most significant factor that depends on the strength of long-range interaction in solutions. Composition-gradient multi-angle static light scattering (CG-MALS) was used to characterize the second virial coefficient of a typical globular protein, bovine serum albumin (BSA), and a larger macromolecule, cowpea mosaic virus (CPMV). Since BSA molecules are prone to aggregate, a composition identification of the BSA solution is carried out with fast protein liquid chromatography (FPLC) and dynamic light scattering (DLS) measurements. The FPLC and DLS study show that monomer is the dominating species in the BSA solutions studied. Then the second virial coefficient of the BSA solution is measured by CG-MALS as a function of pH and univalent salt (NaCl) strengths. The value of second virial coefficient remains greater than zero (corresponding to predominant repulsive interaction between BSA molecules) throughout the experimental region , and the magnitude has a systematic variation as a function of pH and NaCl strength, which reveals the change of the net charge on the surface of BSA molecule and the resulting variation of the inter-molecular force. The minimum of the second virial coefficient is determined to be 1.13*10-5 ml*mol/g² at pH ∼4.6, identified as the isoelectric point of BSA, where the electrostatic interaction reaches the minimum value and van der Waals-London dispersion interaction governs the long-range ordering of the proteins. The second virial coefficient of native CPMV and PEGy (open full item for complete abstract)

Doctor of Education (EdD), Ohio University, 2014, Educational Administration (Education)

This study of Ohio public, private, charter, and vocational school principals during the 2012-2013 school year explored existence or no existence of parental aggression towards school principals. The study researched predictors of aggression towards school principals and identified the frequency of their occurrence. The operationalized definition of parental aggression for this study was parental conduct that the school administrator perceives as threatening to their physical, psychological, or professional well-being. Through regression analysis and descriptive statistics, connections between acts of parental aggression and the demographics of the community, principal, and school were found. This study may serve as a platform to more deeply explore issues relating to aggressive parents, as well as, fill gaps in existing literature and provide insight into possible predictors of parental aggression.

MSARCH, University of Cincinnati, 2013, Design, Architecture, Art and Planning: Architecture

In the past, bazaars constituted a chain of shops along a street that were known to sell a variety of goods. Shopping malls evolved from the traditional form of bazaars where people would frequent to buy goods from local artisans, craftsmen as well as farm products from local farmers. Owing to the oil boom in the Middle East in the late 20th century, particularly, from 1974 - 1980, there was a rapid development of these facilities as people invested oil money in land purchase and construction of commercial buildings. As time went by, investors sought to incorporate leisure facilities into the mall structures thereby creating the modern day malls and bazaars. This was particularly necessary due to the lack of nature parks and other recreational areas, aspects that can be attributed to dry conditions of the region hence limited productive acreage. As a result, malls in Saudi Arabia have become cultural hotspot where people of all ages, nationalities, and races come to meet and interact. The number of people visiting the country has been on the increase. The factors behind visiting this wonderful country are diverse and not limited to the scope of cultural factors and religion, age, gender, among other demographic characteristics. The influence on individuals' capacity to visit shopping malls to purchase their needs, do window shopping, or socialize is therefore varied. The rationale behind this is that while some shoppers seek social exploration, others are after recreational facilities and opportunities. Being an activity that involves face-to-face interactions between buyers and sellers in the mall, shopping has become an agent for breaking cultural boundaries in the society of Saudi Arabia as people interact with each other. This thesis is an observation of the emerging social interactions in the malls in Saudi Arabia.

Doctor of Philosophy, University of Toledo, 2008, Chemistry

DNA replication and repair is one of the most important cellular processes, since preserving the integrity of the DNA genome is essential to all forms of life. Many proteins are involved in the DNA replication process, and their interaction ensures that the DNA is duplicated and repaired in a coordinated and efficient manner. Bacteriophage T4 is a very good model to study DNA replication, since it encodes all the proteins required at the replication fork, proteins which have been extensively characterized. However, how these proteins interact and coordinate the replication process is still largely unknown. One of these interactions that appears to govern the rate and efficiency of the lagging strand synthesis occurs between the 5' to 3' exonuclease RNase H and the single-stranded DNA-binding 32 protein. The interaction between these two proteins is the focus of this work. RNase H and the 32 protein, as well as a number of mutants and truncations, were cloned, expressed and purified. These proteins were then used to form different variants of the RNase H + 32 protein complex, which were characterized through biophysical and structural studies. A crystal structure was obtained for the RNase H + 32-B truncation. This structure, along with the results obtained from the biophysical experiments, provides valuable information on how these two proteins interact to coordinate the lagging strand DNA replication. c Finally, the study of the interaction between RNase H and the 32 protein from bacteriophage Rb 69, a phage related to bacteriophage T4, was also initiated.

Master of Arts, The Ohio State University, 2011, EDU Teaching and Learning

Viewing the goal of language teaching as preparing learners to communicate successfully in the languages and cultures being studied, this thesis adopts the Performed Culture Approach, which expands the meaning of “culture” and integrates the acquisition of behavioral culture with the learning of the target language. This study addresses in-school behavioral culture as one of the primary and fundamental microcultures in language teaching. The assumption is that since schooling is considered a process of socialization and enculturation, the behavior patterns and social norms in teacher-student and student-student interactions have implications for language teaching and learning. With this assumption, this study conducted a questionnaire survey among college students and teachers in American and Chinese cultures. The purpose of the survey is to explore how students of the two cultures behave in the perceived course of transactions with their teachers and classmates, and what the expectation of the teachers in the two cultures are with respect to how students in the teachers' culture are expected to behave in these transactions. Five transactions were examined in the survey: greeting one's teacher, expressing different opinions from one's teacher, dealing with a problem in a study group, responding to compliments from other classmates, and returning a favor to a classmate. Comparisons between the results of the American student respondents and those of the Chinese student respondents indicate differences in terms of the perception the student respondents of the two cultures have as to how they would do things in these transactions. The results of the teacher survey reveal that the ways that the students from the two cultures would be most likely to behave were recognized and accepted by members of their own culture. In addition, the results of the survey virtually confirm the researcher's preliminary observations. This indicates that the perceived course of these trans (open full item for complete abstract)

Doctor of Philosophy, The Ohio State University, 2011, Chemical and Biomolecular Engineering

For bulk production of high purity protein drugs to meet FDA standards, pharmaceutical industries typically process proteins by repeated precipitations and crystallizations. The solution conditions for protein phase separation are routinely determined by trial and error. A thermodynamic model of underlying weak protein-protein interactions will help in a rapid screening of solution conditions in obtaining protein crystals of desired quality and stability. The principal objective of the thesis is, therefore, to understand the fundamental molecular level interactions among different components of protein solutions: protein, water and co-solvents, in particular, the role of hydration and co-solvent preferential interactions on protein-protein interactions. Spatial heterogeneities in protein chemistry and surface topography results in uneven specific hydration of protein surface, which alters the protein-protein interactions by eliminating some complimentary configurations. The dynamics of water at such specific hydration sites was examined in terms of average water residence times and average vacancy times and found to have little impact on protein-protein interactions. The influence of local heterogeneities in surface charge and surface roughness on specific hydration and water dynamics have also been examined. A detailed investigation of the effect of surface curvature on hydration revealed that hydration of a concave surface is thermodynamically expensive than the hydration of a chemically equivalent convex surface. The concave surface is found to remain hydrated only when the interaction between the water and constituent surface atoms are attractive. Protein phase separation is typically induced by adding a precipitating agent or co-solvent, such as inorganic salts or organic compounds: alcohols, polyols, or polyethylene glycol (PEG). Addition of a cosolvent to protein solutions alter the preferential hydration of proteins depending upon its affinity to interact (open full item for complete abstract)

Doctor of Philosophy, The Ohio State University, 2009, Chemistry

Protein–protein interactions (PPIs) play a key role in most biological processes. Many of these interactions are necessary for cell survival. To understand the molecular mechanisms of biological processes, it is essential to study and characterize protein-protein interactions, identify interacting partners and protein interaction networks. There are a number of methods that have been developed to study protein-protein interactions in vitro and in vivo, such as yeast-2-hybrid, fluorescence resonance energy transfer, co-immunoprecipitation, etc. Split protein reassembly is an in vivo probe of protein interactions that circumvents some of the problems with yeast 2-hybrid (indirect interactions, false positives) and co-immunoprecipitation (loss of weak and transient interactions, decompartmentalization). Split GFP reassembly is especially attractive because the GFP chromophore forms spontaneously on protein folding in almost every cell type. However, existing split systems have limitations of evolving cellular fluorescence slowly (3-4 days), failure to evolve at all for some interactions, and also failure to work at a physiological temperature. Among different variants of GFP tested, we found that split folding-reporter GFP (frGFP, a hybrid of EGFP and GFPuv) evolves fluorescence much faster (24 - 30 h) with associating peptides and also evolves fluorescence for the RING domain BRCA1/BARD1 wild type pair. Thirty six known cancer-associated BRCA1 RING domain mutants were tested with split-frGFP system for their role in BRCA1/BARD1 interactions. Some of these mutations resulted in significant reduction of complex reassembly and cellular fluorescence.Split frGFP fragments were further improved by directed evolution (error-prone PCR and DNA shuffling) to obtain fragments for fast and efficient fluorescence reassembly. The evolved fragments were able to generate fluorescence in as little as 12-16 h at 30 °C and in 10-14 h at 37 °C. This system was successfully tested for the (open full item for complete abstract)