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Synthesis Of Gadolinium-Doxorubicin Prodrug Carrying Functional Nanoceria For The Targeted Drug Delivery And Cancer Treatment, Arth Patel
Electronic Theses & Dissertations
The main focus of this research was the development of a polymer-coated nanoceria (PNC) platform to be used as a drug delivery system. Water-dispersible PNC is synthesized using a water-based alkaline precipitation method. Cerium nitrate hexahydrate and poly (acrylic acid) are used for the preparation of PNC. The synthesized PNC was characterized using ZETA, and UV- Vis characterization techniques. Polyacrylic acid (PAA)-coated cerium oxide nanoparticles fabricated for the targeted combination therapy of TNBC (MDA-MB-231) and MCF-7. Using EDC/NHS chemistry, the surface carboxylic acid groups of nanoceria was designed and synthesized with ICAM-1 antibody to target ICAM-1 overexpressing TNBC. Next, doxorubicin …
Regional Gene Repression By Dna Double-Strand Breaks In G1 Phase Cells, Caitlin E. Purman, Patrick L. Collins, Sofia I. Porter, Ankita Saini, Harshath Gupta, Barry P. Sleckman, Eugene M. Oltz
Regional Gene Repression By Dna Double-Strand Breaks In G1 Phase Cells, Caitlin E. Purman, Patrick L. Collins, Sofia I. Porter, Ankita Saini, Harshath Gupta, Barry P. Sleckman, Eugene M. Oltz
Open Access Publications
DNA damage responses (DDR) to double-strand breaks (DSBs) alter cellular transcription programs at the genome-wide level. Through processes that are less well understood, DSBs also alter transcriptional responses locally, which may be important for efficient DSB repair. Here, we developed an approach to elucidate the
In Silico Sequence Optimization For The Reproducible Generation Of Dna Structures, Michael D. Tobiason
In Silico Sequence Optimization For The Reproducible Generation Of Dna Structures, Michael D. Tobiason
Boise State University Theses and Dissertations
Biologically, deoxyribonucleic acid (DNA) molecules have been used for information storage for more than 3 billion years. Today, modern synthesis tools have made it possible to use synthetic DNA molecules as a material for engineering nanoscale structures. These self-assembling structures are capable of both resolutions as fine as 4 angstroms and executing programed dynamic behavior. Numerous approaches for creating structures from DNA have been proposed and validated, however it remains commonplace for engineered systems to exhibit unexpected behaviors such as low formation yields, poor performance, or total failure. It is plausible that at least some of these behaviors arise due …
Nanoscale Optical And Correlative Microscopies For Quantitative Characterization Of Dna Nanostructures, Christopher Michael Green
Nanoscale Optical And Correlative Microscopies For Quantitative Characterization Of Dna Nanostructures, Christopher Michael Green
Boise State University Theses and Dissertations
Methods to engineer nanomaterials and devices with uniquely tailored properties are highly sought after in fields such as manufacturing, medicine, energy, and the environment. The macromolecule deoxyribonucleic acid (DNA) enables programmable self-assembly of nanostructures with near arbitrary shape and size and with unprecedented precision and accuracy. Additionally, DNA can be chemically modified to attach molecules and nanoparticles, providing a means to organize active materials into devices with unique or enhanced properties. One particularly powerful form of DNA-based self-assembly, DNA origami, provides robust structures with the potential for nanometer-scale resolution of addressable sites. DNA origami are assembled from one large DNA …
Effects Of The Dihydrouracil Lesion On Dna Using 1h/31p 1d And 2d Solution Nmr, Benjamin M. Boyd
Effects Of The Dihydrouracil Lesion On Dna Using 1h/31p 1d And 2d Solution Nmr, Benjamin M. Boyd
MSU Graduate Theses
The effects of the dihydrouracil lesion in DNA were studied using two dimensional NMR spectroscopy. The sequence used was based off of the Drew-Dickerson Dodecamer, with the cytosine in the three position replaced by a dihydrouracil. All of the nonexchangeable proton chemical shifts, with the exception of the H2, H5’, and H5’’, of the lesioned DNA were identified using NOESY spectra and then compared to the chemical shift values of the Drew Dickerson Dodecamer. The largest differences in chemical shifts were observed in the nucleotides neighboring the lesion, both within the strand and on the opposite strand. The imino exchangeable …
Identification And Molecular Analysis Of Dna In Exosomes, Jena Tavormina
Identification And Molecular Analysis Of Dna In Exosomes, Jena Tavormina
Dissertations & Theses (Open Access)
Exosomes are heterogeneous nanoparticles 50-150nm in diameter. Exosomes contain many functional cargo components, such as protein, DNA, and RNA. While protein and RNA exosome content has been extensively studied, very little work has been done to characterize exosomal DNA. Here, we demonstrate that exosomal DNA is heterogeneous and its packaging into exosomes is dependent on the cell of origin. Furthermore, through a rigorous assessment of various isolation methods, we identify Size Exclusion Chromatography (SEC) as the best method for the isolation of exosomal DNA for downstream applications. Additionally, we evaluate the methylation status of exosomal DNA and demonstrate that exosomal …
Dna Mixture Statistics Using A Likelihood Ratio Software Tool: Effect Of Variations In Drop-Out Rates And Number Of Contributors, Felicia M. Lucero
Dna Mixture Statistics Using A Likelihood Ratio Software Tool: Effect Of Variations In Drop-Out Rates And Number Of Contributors, Felicia M. Lucero
Student Theses
Complex DNA mixtures can be very probative evidence, but comparisons to a person of interest can be affected by allelic drop-out and uncertainty regarding the number of individuals having contributed DNA to a sample. Scientific organizations such as the International Society of Forensic Genetics (Gill et al., 2006) recommend that likelihood ratios should be used to provide a statistical weight when a positive association is made between the DNA profile of a person of interest and an evidentiary DNA sample. To this effect the New York City Office of Chief Medical Examiner (OCME) developed a software program, Forensic Statistical Tool …
Catching Killers With Consumer Genetic Information, Angela Hackstadt
Catching Killers With Consumer Genetic Information, Angela Hackstadt
University Libraries Faculty Scholarship
In April 2018, Joseph James D'Angelo was arrested as a suspect in the Golden State Killer case. DNA evidence collected at a 1980 crime scene finally shed light on the murderer's identity in early 2018 when investigators turned to GEDMatch, a service that allows users to upload and share DNA data obtained from consumer genetic tests. Consumer genetic testing, DNA collection, and familial DNA searching all raise ethical and privacy concerns. If investigators are using genetic genealogy to solve cold cases, where does that leave consumers?
A New Algorithm For Primer Design, Debanjan Guha Roy
A New Algorithm For Primer Design, Debanjan Guha Roy
Electronic Thesis and Dissertation Repository
The Polymerase Chain Reaction (PCR) technology is widely used to create DNA copies. It has impacted many diverse fields including genetics, forensics, molecular paleontology, medical applications and environmental microbiology.
The main object in PCR is a primer, a short single strand of DNA, about 18-25 bases long, that serves as the starting point of DNA synthesis. Primers are essential for DNA replication because the enzymes that catalyze this process, DNA polymerases, can only add new nucleotides to an existing strand of DNA. The PCR starts at the 3’-end of the primer and copies the opposite strand.Designing good primers is essential …
Embryonic Origin And Genetic Basis Of Cave Associated Phenotypes In The Isopod Crustacean Asellus Aquaticus., Hafasa Mojaddidi, Franco E Fernandez, Priscilla A Erickson, Meredith E. Protas
Embryonic Origin And Genetic Basis Of Cave Associated Phenotypes In The Isopod Crustacean Asellus Aquaticus., Hafasa Mojaddidi, Franco E Fernandez, Priscilla A Erickson, Meredith E. Protas
Meredith Protas
Characteristics common to animals living in subterranean environments include the reduction or absence of eyes, lessened pigmentation and enhanced sensory systems. How these characteristics have evolved is poorly understood for the majority of cave dwelling species. In order to understand the evolution of these changes, this study uses an invertebrate model system, the freshwater isopod crustacean, Asellus aquaticus, to examine whether adult differences between cave and surface dwelling individuals first appear during embryonic development. We hypothesized that antennal elaboration, as well as eye reduction and pigment loss, would be apparent during embryonic development. We found that differences in pigmentation, eye …
Gut Dna Virome Diversity And Its Association With Host Bacteria Regulate Inflammatory Phenotype And Neuronal Immunotoxicity In Experimental Gulf War Illness, Ratanesh K. Seth, Rabia Maqsood, Ayan Mondal, Dipro Bose, Diana Kimono, Larinda A. Holland, Patricia Janulewicz Lloyd, Nancy Klimas, Ronnie Horner, Kimberly Sullivan, Efrem S. Lim, Saurabh Chatterjee
Gut Dna Virome Diversity And Its Association With Host Bacteria Regulate Inflammatory Phenotype And Neuronal Immunotoxicity In Experimental Gulf War Illness, Ratanesh K. Seth, Rabia Maqsood, Ayan Mondal, Dipro Bose, Diana Kimono, Larinda A. Holland, Patricia Janulewicz Lloyd, Nancy Klimas, Ronnie Horner, Kimberly Sullivan, Efrem S. Lim, Saurabh Chatterjee
Faculty Publications
Gulf War illness (GWI) is characterized by the persistence of inflammatory bowel disease, chronic fatigue, neuroinflammation, headache, cognitive impairment, and other medically unexplained conditions. Results using a murine model show that enteric viral populations especially bacteriophages were altered in GWI. The increased viral richness and alpha diversity correlated positively with gut bacterial dysbiosis and proinflammatory cytokines. Altered virome signature in GWI mice also had a concomitant weakening of intestinal epithelial tight junctions with a significant increase in Claudin-2 protein expression and decrease in ZO1 and Occludin mRNA expression. The altered virome signature in GWI, decreased tight junction protein level was …
Managing Microbial Contamination Of Surface Water Using Ozone, Shannon Deehan
Managing Microbial Contamination Of Surface Water Using Ozone, Shannon Deehan
Electronic Theses and Dissertations
The increasing levels of fecal pollution in recreational waters is becoming a major concern since it poses a health risk and has negative impacts on aquatic life and the economy. The major threats to water quality come from point sources such as combined sewer overflows (CSO) and sanitary sewer overflows (SSO). The most sustainable solution is to stop the problem at the source, therefore, treatment of the overflows using ozonation is the subject of study in this thesis. Culture-dependent methods (IDEXX Colilert) are currently being used by regulatory agencies but due to several disadvantages of these methods culture-independent methods are …
Understanding Evolutionary Relationships Between Sedges Genus Rhynocospora, Abidell T. Thomas
Understanding Evolutionary Relationships Between Sedges Genus Rhynocospora, Abidell T. Thomas
Honors Theses
The relationships among species in the genus Rhyncospora, a member of the flowering plant family Cyperaceae, are of value to study as they are essential plant components to our ecosystem. Sedges are necessary for wetland preservation, land-sea barriers, and in many cases serve as a natural water detoxifier. Rhyncospora is a prime genus to ask questions about their phylogeny because their evolutionary history is poorly understood for them to be so prevalent. To understand the relationships between a specific group of sedges (genus Rhyncospora) I utilized nucleotide sequence data in a phylogenetic approach. Phylogenetics is the study of …
Evidence For P53-Mediated Induction Of Wrap53a In Response To Dna Damage, Anne Shelton Hucks
Evidence For P53-Mediated Induction Of Wrap53a In Response To Dna Damage, Anne Shelton Hucks
Theses and Dissertations
p53 is a powerful tumor suppressor mutated in approximately half of all cancers. Its mRNA is stabilized post-transcriptionally via complementary base pairing with the transcript of its antisense gene, WRAP53α; without this interaction, p53 protein cannot accumulate enough to carry out its many functions related to apoptosis, cell cycle arrest, and DNA damage repair. Previous studies have shown that WRAP53α is induced in response to DNA damage. The purpose of this study was to determine which transcription factors might be responsible for this induction. After identifying three putative p53 binding sites on the WRAP53α promoter, we used chromatin immunoprecipitation …
Integrated Transcriptome Analysis Reveals Mirna-Mrna Crosstalk In Laryngeal Squamous Cell Carcinoma., Yang Zhang, Yong Chen, Jinhai Yu, Guiming Liu, Zhigang Huang
Integrated Transcriptome Analysis Reveals Mirna-Mrna Crosstalk In Laryngeal Squamous Cell Carcinoma., Yang Zhang, Yong Chen, Jinhai Yu, Guiming Liu, Zhigang Huang
Yong Chen
Next generation sequencing (NGS) has proven to be a powerful tool in delineating myriads of molecular subtypes of cancer, as well as in revealing accumulation of genomic mutations throughout cancer progression. Whole genome microRNA (miRNA) and mRNA expression profiles were obtained from patients with laryngeal squamous cell carcinoma (LSCC) using deep sequencing technology, and were analyzed by utilizing integrative computational approaches. A large number of protein-coding and non-coding genes were detected to be differentially expressed, indicating a functional switch in LSCC cells. A total of 127 mutated genes were detected to be significantly associated with ectoderm and epidermis development. Eleven …
Climp: Clustering Motifs Via Maximal Cliques With Parallel Computing Design., Shaoqiang Zhang, Yong Chen
Climp: Clustering Motifs Via Maximal Cliques With Parallel Computing Design., Shaoqiang Zhang, Yong Chen
Yong Chen
A set of conserved binding sites recognized by a transcription factor is called a motif, which can be found by many applications of comparative genomics for identifying over-represented segments. Moreover, when numerous putative motifs are predicted from a collection of genome-wide data, their similarity data can be represented as a large graph, where these motifs are connected to one another. However, an efficient clustering algorithm is desired for clustering the motifs that belong to the same groups and separating the motifs that belong to different groups, or even deleting an amount of spurious ones. In this work, a new motif …
Fishermp: Fully Parallel Algorithm For Detecting Combinatorial Motifs From Large Chip-Seq Datasets., Shaoqiang Zhang, Ying Liang, Xiangyun Wang, Zhengchang Su, Yong Chen
Fishermp: Fully Parallel Algorithm For Detecting Combinatorial Motifs From Large Chip-Seq Datasets., Shaoqiang Zhang, Ying Liang, Xiangyun Wang, Zhengchang Su, Yong Chen
Yong Chen
Detecting binding motifs of combinatorial transcription factors (TFs) from chromatin immunoprecipitation sequencing (ChIP-seq) experiments is an important and challenging computational problem for understanding gene regulations. Although a number of motif-finding algorithms have been presented, most are either time consuming or have sub-optimal accuracy for processing large-scale datasets. In this article, we present a fully parallelized algorithm for detecting combinatorial motifs from ChIP-seq datasets by using Fisher combined method and OpenMP parallel design. Large scale validations on both synthetic data and 350 ChIP-seq datasets from the ENCODE database showed that FisherMP has not only super speeds on large datasets, but also …
Haplotype Analysis For Irish Ancestry, Heather Miller Coyle, Robert Whiting
Haplotype Analysis For Irish Ancestry, Heather Miller Coyle, Robert Whiting
Forensic Science Publications
Forensic haplotype analysis of the male Y chromosome is currently used to establish the number of male donors in sexual assaults, the number of male bleeders in blood pattern analysis, and for ancestry correlation to genetic founder populations in biogeographic studies. In forensic laboratory applications, its primary use is for DNA profile generation with trace amounts of male DNA in the presence of excess female DNA (e.g. spermatozoa identification, male component of fingernail scrapings). Our study supports the potential use of the Y chromosome in a “dragnet” approach (most haplotypes are unique) similar to that described by Kayser in 2017 …
Dna Recovery From Handwritten Documents Using A Novel Vacuum Technique, Patrick M. Mclaughlin
Dna Recovery From Handwritten Documents Using A Novel Vacuum Technique, Patrick M. Mclaughlin
Student Theses
Investigations of many crimes such as robberies, kidnappings, and terrorism are often associated with the recovery of a paper document which has been written by the perpetrator. Paper can provide a variety of forensic evidence such as DNA, latent fingermarks, and indented writing. The focus of this study was the collection of probative DNA profiles from the text region of a handwritten note through a vacuum suction device without altering or destroying the document. Collection of DNA evidence was carried out in two separate groups. The first group involved 11 volunteers providing a handwritten note sample with unwashed hands. The …
Engineering Dna Recognition And Allosteric Response Properties Of Tetr Family Proteins By Using A Module-Swapping Strategy, Rey P. Dimas, Benjamin R. Jordan, Xian-Li Jiang, Catherine Martini, Joseph S. Glavy, Dustin P. Patterson, Faruck Morcos, Clement T.Y. Chan
Engineering Dna Recognition And Allosteric Response Properties Of Tetr Family Proteins By Using A Module-Swapping Strategy, Rey P. Dimas, Benjamin R. Jordan, Xian-Li Jiang, Catherine Martini, Joseph S. Glavy, Dustin P. Patterson, Faruck Morcos, Clement T.Y. Chan
Biology Faculty Publications and Presentations
The development of synthetic biological systems requires modular biomolecular components to flexibly alter response pathways. In previous studies, we have established a module-swapping design principle to engineer allosteric response and DNA recognition properties among regulators in the LacI family, in which the engineered regulators served as effective components for implementing new cellular behavior. Here we introduced this protein engineering strategy to two regulators in the TetR family: TetR (UniProt Accession ID: P04483) and MphR (Q9EVJ6). The TetR DNA-binding module and the MphR ligand-binding module were used to create the TetR-MphR. This resulting hybrid regulator possesses DNA-binding properties of TetR and …
That Other Form Of Madness: A Multidisciplinary Study Of Infectious Disease Within The Milwaukee County Poor Farm Cemetery, Helen Marie Werner
That Other Form Of Madness: A Multidisciplinary Study Of Infectious Disease Within The Milwaukee County Poor Farm Cemetery, Helen Marie Werner
Theses and Dissertations
Between the years of 1882 and 1925, the Milwaukee County Poor Farm buried several
thousand members of Milwaukee’s indigent population in what would later be designated
Cemetery II. In 1991 and early 1992, after discovery of the cemetery during construction of parts
of the Milwaukee Regional Medical Center, 1,649 burials were excavated. The graves had long
been abandoned and the headstones bulldozed, leaving a register of burials without any obvious
way of associating each individual with their identity. A copy of the register is curated at the
University of Wisconsin-Milwaukee Archaeological Research Laboratory. The Milwaukee
County Poor Farm was a …
Copper And Barium As Dietary Discriminants: The Effects Of Diagenesis, J B. Lambert, S V. Simpson, Carole B. Szpunar, J E. Buikstra
Copper And Barium As Dietary Discriminants: The Effects Of Diagenesis, J B. Lambert, S V. Simpson, Carole B. Szpunar, J E. Buikstra
Carole Szpunar
No abstract provided.
Enhancement Aes Based On 3d Chaos Theory And Dna Operations Addition, Alaa Kadhim, Rasha Subhi Ali
Enhancement Aes Based On 3d Chaos Theory And Dna Operations Addition, Alaa Kadhim, Rasha Subhi Ali
Karbala International Journal of Modern Science
The techniques modern used to encrypt text data play an important role in preventing unauthorized access. The AES algorithm is widely used in various cryptographic applications, but coz the algorithm has been subjected to several successive attacks, the need is necessary to improve the algorithm and make it more powerful the algorithm.In dis search will be inserted plaintext 1024 will be divided into four blocks each Block consists of 256 bits and each Block will pass the four stages represent(substitution,shift,Mix column and round key).A new step will be added to the four stages of permutation. Where chaos theory is adopted …
Computational Analysis And Design Optimization Of Convective Pcr Devices, Jung Il Shu
Computational Analysis And Design Optimization Of Convective Pcr Devices, Jung Il Shu
Mechanical & Aerospace Engineering Theses & Dissertations
Polymerase Chain Reaction (PCR) is a relatively novel technique to amplify a few copies of DNA to a detectable level. PCR has already become common in biomedical research, criminal forensics, molecular archaeology, and so on. Many have attempted to develop PCR devices in numerous types for the purpose of the lab-on-chip (LOC) or point-of-care (POC). To use PCR devices for POC lab testing, the price must be lower, and the performance should be comparable to the lab devices. For current practices with the existing methods, the price is pushed up higher partially due to too much dependence on numerous developmental …
The Signature Of God In Medicine And Microbiology An Apologetic Argument For Declarative Design In The Discoveries Of Alexander Fleming, Alan L. Gillen, Michael Cargill
The Signature Of God In Medicine And Microbiology An Apologetic Argument For Declarative Design In The Discoveries Of Alexander Fleming, Alan L. Gillen, Michael Cargill
Alan L. Gillen
In logic and reasoning, a signature indicates the presence of an author; likewise, the characteristics of staphylococci indicate the presence of a Creator. Staphylococci and its “kind” are common bacteria, particularly in colonized people.1 Staphylococcus aureus has a complex molecular mechanism of assembling its golden pigment, staphyloxanthin. The biosynthesis of staphyloxanthin is a stellar example of irreducible complexity. Similar to staphylococci, the life and works of Alexander Fleming show the fingerprints of Providence. The so-called “serendipitous” achievements of Fleming have contributed to modern medicine, convincing Fleming and others that God was at work in his life. Fleming recognized that …
Unrequited Innocence In U.S. Capital Cases: Unintended Consequences Of The Fourth Kind, Rob Warden, John Seasly
Unrequited Innocence In U.S. Capital Cases: Unintended Consequences Of The Fourth Kind, Rob Warden, John Seasly
Northwestern Journal of Law & Social Policy
No abstract provided.
Fishermp: Fully Parallel Algorithm For Detecting Combinatorial Motifs From Large Chip-Seq Datasets., Shaoqiang Zhang, Ying Liang, Xiangyun Wang, Zhengchang Su, Yong Chen
Fishermp: Fully Parallel Algorithm For Detecting Combinatorial Motifs From Large Chip-Seq Datasets., Shaoqiang Zhang, Ying Liang, Xiangyun Wang, Zhengchang Su, Yong Chen
Faculty Scholarship for the College of Science & Mathematics
Detecting binding motifs of combinatorial transcription factors (TFs) from chromatin immunoprecipitation sequencing (ChIP-seq) experiments is an important and challenging computational problem for understanding gene regulations. Although a number of motif-finding algorithms have been presented, most are either time consuming or have sub-optimal accuracy for processing large-scale datasets. In this article, we present a fully parallelized algorithm for detecting combinatorial motifs from ChIP-seq datasets by using Fisher combined method and OpenMP parallel design. Large scale validations on both synthetic data and 350 ChIP-seq datasets from the ENCODE database showed that FisherMP has not only super speeds on large datasets, but also …
The Law Of Genetic Privacy: Applications, Implications, And Limitations, Ellen Wright Clayton, Barbara J. Evans, James W. Hazel, Mark A. Rothstein
The Law Of Genetic Privacy: Applications, Implications, And Limitations, Ellen Wright Clayton, Barbara J. Evans, James W. Hazel, Mark A. Rothstein
Vanderbilt Law School Faculty Publications
Recent advances in technology have significantly improved the accuracy of genetic testing and analysis, and substantially reduced its cost, resulting in a dramatic increase in the amount of genetic information generated, analysed, shared, and stored by diverse individuals and entities. Given the diversity of actors and their interests, coupled with the wide variety of ways genetic data are held, it has been difficult to develop broadly applicable legal principles for genetic privacy. This article examines the current landscape of genetic privacy to identify the roles that the law does or should play, with a focus on federal statutes and regulations, …
Dna Methods To Understand Genetic Variation Of The Mola Mola Populations In And Near Cape Cod, Jessica Barros
Dna Methods To Understand Genetic Variation Of The Mola Mola Populations In And Near Cape Cod, Jessica Barros
Honors Program Theses and Projects
The ocean sunfish Mola mola is a frequent summer visitor to the waters off of Southeastern Massachusetts and Cape Cod. In late fall, water temperatures drop and some animals strand on area beaches. Stranded animals provide an easily available source of DNA to study the population genetics of this species with the specific goal to learn about genetic variation within and between individuals in this population. This study will continue to identify and characterize microsatellite repeat regions that will be used to study genetic variation. Four new microsatellite regions have been identified and amplified using polymerase chain reaction conditions that …
Experimental Evidence Supportive Of The Quantum Dna Model, F. Matthew Mihelic
Experimental Evidence Supportive Of The Quantum Dna Model, F. Matthew Mihelic
Faculty Publications
The DNA molecule can be modeled as a quantum logic processor in which electron spin qubits are held coherently in each nucleotide in a logically and thermodynamically reversible enantiomeric symmetry, and can be coherently conducted along the pi-stacking interactions of aromatic nucleotide bases, while simultaneously being spin-filtered via the helicity of the DNA molecule. Entangled electron pairs can be separated by that spin-filtering, held coherently at biological temperatures in the topologically insulated nucleotide quantum gates, and incorporated into separate DNA strands during DNA replication. Two separate DNA strands that share quantum entangled electrons can be mitotically divided into individual cells, …