Whole Exome Sequencing Identifies A Novel Mutation In Aspm And Ultra-Rare Mutation In Cdk5rap2 Causing Primary Microcephaly In Consanguineous Pakistani Families,
2022
Government College University, 38000, Faisalabad, Pakistan
Whole Exome Sequencing Identifies A Novel Mutation In Aspm And Ultra-Rare Mutation In Cdk5rap2 Causing Primary Microcephaly In Consanguineous Pakistani Families, Ehtisham Ul Haq Makhdoom, Haseeb Anwar, Shahid Mahmood Baig, Ghulam Hussain
Department of Biological & Biomedical Sciences
Background & objectives: Primary Microcephaly (MCPH) is a rare neurogenetic disease, manifesting congenitally reduced head circumference and non-progressive intellectual disability (ID). To date, twenty-eight genes with biallelic mutations have been reported for this disorder. The study aimed for molecular genetic characterization of Pakistani families segregating MCPH.
Methods: We studied two unrelated consanguineous families (family A and B) presenting >2 patients with diagnostic symptoms of MCPH, born to asymptomatic parents. We employed whole-exome sequencing (WES) of probands to find putative causal mutations. The candidate variants were further confirmed and analyzed for co-segregation by Sanger sequencing of all available members of each …
Introduction Of Plasmid Dna Into Sneathia Vaginalis; The First Step To Genetic Manipulation,
2022
Virginia Commonwealth University
Introduction Of Plasmid Dna Into Sneathia Vaginalis; The First Step To Genetic Manipulation, Rea Mansukhani, Cathyrn O'Brien, Zion Mccoy, Kimberly K. Jefferson
UROP Posters
Many bacterial pathogens are capable of secreting one or more toxins, all of which can be categorized based on their function, target, or mechanism of action. Pore-forming toxins, for instance, are characterized based on their ability to perforate the host membrane. This may result in the delivery of bacterial substances into the host cell, release of compounds from the host cell, and/or death of the host cell. Research from our lab has demonstrated that Sneathia vaginalis, a gram-negative anaerobic bacterial species that is associated with bacterial vaginosis and preterm birth, produces a pore-forming toxin referred to as cytopathogenic toxin …
Transitions In Bacterial Communities Across Two Fermentation-Based Virgin Coconut Oil (Vco) Production Processes,
2022
Ateneo de Manila University
Transitions In Bacterial Communities Across Two Fermentation-Based Virgin Coconut Oil (Vco) Production Processes, Zomesh A N Maini, Crisanto M. Lopez
Biology Faculty Publications
Despite being one of the most used methods of virgin coconut oil (VCO) production, there is no metagenomic study that details the bacterial community shifts during fermentation-based VCO production. The identification and quantification of bacteria associated with coconut milk fermentation is useful for detecting the dominant microbial genera actively involved in VCO production which remains largely undescribed. Describing the constitutive microbial genera involved in this traditional fermentation practice can be used as a preliminary basis for improving industrial practices and developing better fermentation procedures. In this study, we utilized 16S rRNA metagenomic sequencing to trace the transitions in microbial community …
Antibacterial Activity Of Commiphora Molmol (Myrrha) Against The Periodontal Pathogen, Aggregatibacter Actinomycetemcomitans,
2022
Long Island University
Antibacterial Activity Of Commiphora Molmol (Myrrha) Against The Periodontal Pathogen, Aggregatibacter Actinomycetemcomitans, Khalid Nafea Alharbi
Selected Full-Text Master Theses 2021-
The exogenous pathogen Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) is most frequently involved in periodontitis and other systemic diseases. The treatment of these infections involves antibiotic therapy with Amoxicillin being the most popular antibiotic against A. actinomycetemcomitans but now, the drug is not 100% effective due to the development of 0-84% antibiotic resistance. The present study aimed to determine antibacterial activity of Commiphora molmol (Myrrha) against the periodontal pathogen, Aggregatibacter actinomycetemcomitans. In detail, the strain LIU1239 of Aggregatibacter actinomycetemcomitans was used where the plasmid pVJT128 is used for transposon mutagenesis and the Escherichia coli strain MV10Nal (stock # LIU4). The …
Phip Variants Associated With Chung–Jansen Syndrome Disrupt Replication Fork Stability And Genome Integrity,
2022
University of South Florida
Phip Variants Associated With Chung–Jansen Syndrome Disrupt Replication Fork Stability And Genome Integrity, Neysha Tirado-Class, Caitlin Hathaway, Wendy K. Chung, Huzefa Dungrawala
Molecular Biosciences Faculty Publications
Chung–Jansen syndrome (CJS) is a rare, autosomal dominant disorder characterized by developmental delay, intellectual disability/cognitive impairment, behavioral challenges, obesity, and dysmorphic features. CJS is associated with heterozygous variants in PHIP (Pleckstrin-Homology Interacting Protein), a gene that encodes one of several substrate receptors for Cullin4-RING (CRL4) E3 ubiquitin ligase complex. Full-length PHIP, also called DCAF14, was recently identified to function as a replication stress response protein. Herein, we report the identification of two PHIP missense variants identified by exome sequencing in unrelated individuals with CJS. The variants p.D488V and p.E963G occur in different functional elements of DCAF14-WD40 repeat domain and pleckstrin …
Sequence Properties Of An Intramolecular Interaction That Inhibits P53 Dna Binding,
2022
University of South Florida
Sequence Properties Of An Intramolecular Interaction That Inhibits P53 Dna Binding, Emily Gregory, Gary W. Daughdrill
Molecular Biosciences Faculty Publications
An intramolecular interaction between the p53 transactivation and DNA binding domains inhibits DNA binding. To study this autoinhibition, we used a fragment of p53, referred to as ND WT, containing the N-terminal transactivation domains (TAD1 and TAD2), a proline rich region (PRR), and the DNA binding domain (DBD). We mutated acidic, nonpolar, and aromatic amino acids in TAD2 to disrupt the interaction with DBD and measured the effects on DNA binding affinity at different ionic strengths using fluorescence anisotropy. We observed a large increase in DNA binding affinity for the mutants consistent with reduced autoinhibition. The ΔΔG between DBD and …
Isolation And Characterization Of Novel B. Subtilis Bacteriophages,
2022
Belmont University
Isolation And Characterization Of Novel B. Subtilis Bacteriophages, Johnson Phung, Rebecca Adams
Science University Research Symposium (SURS)
Bacteriophages are viruses that infect bacteria cells. Bacteria, being the most abundant form of life on the planet, have an estimated ten to a hundred different types of bacteriophages that infect each species. Bacteriophages are everywhere on Earth, but the number of phages that have been identified is only a fraction of what are thought to exist. This project focuses on isolating and identifying phages that infect the bacterium Bacillus subtilis. B subtilis is a decomposition microbe commonly found in the soil. Our first goal was to determine whether the 3 samples of Bacillus subtilis phages that were previously …
Adenoviral-Vectored Centralized Consensus Hemagglutinin
Vaccine Provides Broad Protection Against H2 Influenza A Virus,
2022
University of Nebraska-Lincoln
Adenoviral-Vectored Centralized Consensus Hemagglutinin Vaccine Provides Broad Protection Against H2 Influenza A Virus, Erika M. Petro-Turnquist, Brianna L. Bullard, Matthew J. Pekarek, Eric A. Weaver
Nebraska Center for Virology: Faculty Publications
Several influenza pandemics have occurred in the past century, one of which emerged in 1957 from a zoonotic transmission of H2N2 from an avian reservoir into humans. This pandemic caused 2–4 million deaths and circulated until 1968. Since the disappearance of H2N2 from human populations, there has been waning immunity against H2, and this subtype is not currently incorporated into seasonal vaccines. However, H2 influenza remains a pandemic threat due to consistent circulation in avian reservoirs. Here, we describe a method of pandemic preparedness by creating an adenoviral-vectored centralized consensus vaccine design against human H2 influenza. We also assessed the …
Computational Strain Analysis In Microbiome Datasets With Application To Skin Disease,
2022
University of Central Florida
Computational Strain Analysis In Microbiome Datasets With Application To Skin Disease, Minerva Fatimae Ventolero
Electronic Theses and Dissertations, 2020-2023
Identifying bacterial strains is important not only for human health research but also for various environmental and industrial applications. The plethora of available shotgun metagenomic datasets provide an unprecedented opportunity to discover novel bacterial strains in different environmental niches. In this thesis, we evaluated the existing novel-strain-based tools and showed that their performance is still suboptimal. Due to the difficulty in distinguishing strains of similar abundance by available tools, the SMS (strains in multiple samples) tool was developed to de novo identify bacterial strains in multiple shotgun metagenomic samples. We showed that SMS distinguishes strains of similar abundance well and …
Elucidation Of Novel And Established Campylobacter Species With Clinical And Agricultural Significance Through Phenotypic, Genotypic, And Taxonomic Investigation,
2022
Department of Biological Sciences, Munster Technological University, Cork, Ireland
Elucidation Of Novel And Established Campylobacter Species With Clinical And Agricultural Significance Through Phenotypic, Genotypic, And Taxonomic Investigation, Caoimhe Lynch
Theses
Campylobacter spp. are well-established human, veterinary and economic pathogens, with a broad host range spanning from terrestrial and marine mammalian, avian and reptilian hosts. The scope of the study includes novel and notorious species within the genus, with reference to zoonotic agents Campylobacter jejuni and Campylobacter coli - the leading cause of human bacterial gastroenteritis in the EU and Ireland, Campylobacter fetus that represents the most common campylobacter causing bacteraemia and two novel species isolated from pigs not described in the literature previously. C. jejuni and C. coli isolates (n = 350) recovered from broiler caecal and neck skin samples, …
Investigating The Development Of Alcohol Tolerance By The Opportunistic Pathogen Staphylococcus Aureus,
2022
West Chester University of Pennsylvania
Investigating The Development Of Alcohol Tolerance By The Opportunistic Pathogen Staphylococcus Aureus, Vaughn Perveiler
West Chester University Master’s Theses
With the ongoing COVID-19 pandemic, the use of alcohol-based antiseptics by the general public has greatly increased. While this form of antimicrobial is effective in stopping the spread of both bacterial and viral infections, little research has been conducted in the development of alcohol tolerance by microorganisms or its mechanisms. In this study, experimental evolution was conducted on four separate strains of Staphylococcus aureus to determine if tolerance can develop towards moderate to high concentrations of ethanol. Once ethanol tolerance was established in the evolved populations, whole genome sequencing was performed to identify the potential mutations responsible for the observed …
Evaluation Of Oasl And Herc5’S Role In The Non-Lytic Clearance Of Influenza A Virus From Club Cells,
2022
West Chester University
Evaluation Of Oasl And Herc5’S Role In The Non-Lytic Clearance Of Influenza A Virus From Club Cells, Steve Crisafulli
West Chester University Master’s Theses
Influenza A virus (IAV) is a highly infectious pathogen responsible for causing severe respiratory illness and death in humans and animals worldwide. Due to highly effective strategies to negate host antiviral defenses, IAV leads to the death of nearly all infected cells. Furthermore, IAV induces high levels of genome-damaging oxidative stress within infected cells and suppresses the cellular mismatch repair (MMR) mechanism, thereby inhibiting expression of key antiviral genes, which further contributes to cell death. However, recent studies have demonstrated that a subset of respiratory epithelial cells, called club cells, are able to non-lytically clear IAV and continue to survive …
Effect Of Metal-Containing Nanoparticles On Bacterial Biofilms And On The Microbiome Of Girardia Tigrina,
2022
West Chester University
Effect Of Metal-Containing Nanoparticles On Bacterial Biofilms And On The Microbiome Of Girardia Tigrina, Azar Saikali
West Chester University Master’s Theses
Anthropogenically-produced nanoparticles are a form of nanotechnology being used in industries including food and textiles. Humans and livestock are frequently exposed to metal-containing nanoparticles (MCNPs), that have been washed into streams and rivers, have been deliberately used in food packaging as antimicrobials, preservatives or for supplementation. The animal microbiome, which consists of a diverse community of microorganisms, provides a number of benefits to the host in terms of nutrition availability, immune support, and can influence behavior. Biofilms of diverse microbes may cause detrimental effects, for instance by causing dental diseases in humans. However, the scientific community has not reached consensus …
Unique Features Of Alarmone Metabolism In Clostridioides Difficile,
2022
Old Dominion University
Unique Features Of Alarmone Metabolism In Clostridioides Difficile, Asia Poudel, Astha Pokhrel, Adenrele Oludiran, Estevan J. Coronado, Kwincy Alleyne, Marrett M. Gilfus, Raj K. Gurung, Surya B. Adhikari, Erin B. Purcell
Chemistry & Biochemistry Faculty Publications
The “magic spot” alarmones (pp)pGpp, previously implicated in Clostridioides difficile antibiotic survival, are synthesized by the RelA-SpoT homolog (RSH) of C. difficile (RSHCd) and RelQCd. These enzymes are transcriptionally activated by diverse environmental stresses. RSHCd has previously been reported to synthesize ppGpp, but in this study, we found that both clostridial enzymes exclusively synthesize pGpp. While direct synthesis of pGpp from a GMP substrate, and (p)ppGpp hydrolysis into pGpp by NUDIX hydrolases, have previously been reported, there is no precedent for a bacterium synthesizing pGpp exclusively. Hydrolysis of the 5′ phosphate or pyrophosphate from GDP …
Copper(Ii) And Silver(I)‑1,10‑Phenanthroline‑5,6‑Dione Complexes Interact With Double‑Stranded Dna: Further Evidence Of Their Apparent Multi‑Modal Activity Towards Pseudomonas Aeruginosa,
2022
Universidade Federal do Rio de Janeiro
Copper(Ii) And Silver(I)‑1,10‑Phenanthroline‑5,6‑Dione Complexes Interact With Double‑Stranded Dna: Further Evidence Of Their Apparent Multi‑Modal Activity Towards Pseudomonas Aeruginosa, Anna Clara Milesi Galdino, Lívia Viganor, Matheus Mendonça Pereira, Michael Devereux, Malachy Mccann, Marta Helena Branquinha, Zara Molphy, Sinéad O'Carroll, Conor Bain, Georgia Menounou, Andrew Kellett, André Luis Souza Dos Santos
Articles
Tackling microbial resistance requires continuous efforts for the development of new molecules with novel mechanisms of action and potent antimicrobial activity. Our group has previously identified metal-based compounds, [Ag(1,10-phenanthroline-5,6-dione)2]ClO4 (Ag-phendione) and [Cu(1,10-phenanthroline-5,6-dione)3](ClO4)2.4H2O (Cu-phendione), with efficient antimicrobial action against multidrug-resistant species. Herein, we investigated the ability of Ag-phendione and Cu-phendione to bind with double-stranded DNA using a combination of in silico and in vitro approaches. Molecular docking revealed that both phendione derivatives can interact with the DNA by hydrogen bonding, hydrophobic and electrostatic interactions. Cu-phendione exhibited the highest binding affinity to either major (− 7.9 kcal/mol) or minor (− 7.2 kcal/mol) …
Role Of Host Restriction Factors On Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Replication,
2022
South Dakota State University
Role Of Host Restriction Factors On Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Replication, Pratik Katwal
Electronic Theses and Dissertations
In this study, the role of IFITM3 on PRRSV replication was studied in vitro by expressing exogenous IFITM3 in MARC-145 cells. An average of 31% reduction in PRRSV N protein expression and an average of 5.4 fold decrease in virus titer in the supernatant were observed in IFITM3 overexpressing cells as compared to vector control cells at 24 hours post infection (hpi). Moreover, there was a positive correlation between interferon- induced IFITM3 up-regulation and reduced PRRSV replication. To determine the role of endogenous IFITM3 in PRRSV replication, siRNA induced knockdown of IFITM3 was employed. RT-PCR validated the successful silencing of …
Decoding The Mystery Of Antibiotic Persistence,
2022
South Dakota State University
Decoding The Mystery Of Antibiotic Persistence, Tahmina Hossain
Electronic Theses and Dissertations
This work combines microbiology, molecular biology, Next-Generation Sequencing and system biology approaches to explore the mechanism of antibiotic persistence: a multi-drug tolerant, non-dividing, and metabolically altered state present in a subpopulation of cells due to phenotypic diversity rather than genetic variation (i.e. mutations). Persister can survive lethal antibiotic state and resuscitate after the treatment period is over. They are considered as the major contributing factor behind recurring infections. They also have a high mutation rate, which increases the chances of bacteria gaining antibiotic resistance. The formation of this phenotypic variant (persister) threatens the therapeutic effectiveness of antibiotics and understanding how …
The Bgs 13 Mutant Of Pichia Pastoris And Its Effect On Structural Changes Of The Reporter Protein B-Lactogolbulin,
2022
University of the Pacific
The Bgs 13 Mutant Of Pichia Pastoris And Its Effect On Structural Changes Of The Reporter Protein B-Lactogolbulin, Bushra Irshad
University of the Pacific Theses and Dissertations
Pichia pastoris, a methylotrophic yeast, is an ideal host for recombinant protein expression. It has the capability of performing many eukaryotic post-translational modifications and grows to high cell densities. However, P. pastoris’s secretion properties are not always efficient, and its secretory pathway mechanisms have not been thoroughly elucidated. A previously identified mutant strain, bgs13, was found to efficiently secrete most recombinant proteins tested, raising the possibility that this bgs13 mutant is a universal super secreter and understanding its secretion process is needed. In this study, we used a reporter protein, ?-lactoglobulin (b-LG), to perform structural analysis and comparisons of protein …
Leveraging Queueing Theory To Develop Advanced Synthetic Biological Circuits,
2022
South Dakota State University
Leveraging Queueing Theory To Develop Advanced Synthetic Biological Circuits, Prajakta K. Jadhav
Electronic Theses and Dissertations
The complexity and redundancy in the network topology of the cell hinder our understanding of the natural system, making it challenging to engineer living organisms with the desired functionality. The long-term goal of synthetic biology and bioengineering is to engineer cells to perform specific functions with increased robustness. The robust engineered systems can be easily applied to different species with the same outcome. An effective approach is a bottom-up approach where we tease apart the biological pathways and study them independently. Understanding the underlying mechanisms of biological pathways can be further applied to construct complex biological circuits that function similarly …
Characterizing The Lir Domain Of Abin1 And Identifying Its Role As A Regulator Of Mitophagy,
2022
South Dakota State University
Characterizing The Lir Domain Of Abin1 And Identifying Its Role As A Regulator Of Mitophagy, Andrew Rhiner
Electronic Theses and Dissertations
A20 Inhibitor of NF-κB (ABIN1/TNIP1) is a known regulator of TNFα signaling induced cell death and inflammation. The regulatory activity has been attributed to ABIN1’s recruitment of the ubiquitin editing enzyme TNF-α-induced protein 3 (TNFAIP3/A20) to Receptor-interacting serine/threonine-protein kinase 1 (RIPK1). The regulation of RIPK1 by ABIN1 and A20 relies on a third player, linear ubiquitin chain assembly complex (LUBAC), which is involved in the recruitment of ABIN1 to RIPK1. Loss of LUBAC or ABIN1 is embryonic lethal, but loss of A20 is not embryonic lethal. The embryonic lethality due to loss of ABIN1, paired with the lack of lethality …
