Early Life Socioeconomic Circumstance and Late Life Brain Hyperintensities : A Population Based Cohort Study

Funding: Image acquisition and image analysis for this study was funded by the Alzheimer's Research Trust (now Alzheimer's Research UK). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Acknowledgments The authors would like to thank the participants of the Aberdeen 1936 Birth Cohort (ABC36), without whom this research would not have been possible.Peer reviewedPublisher PD

Preventive behaviors and information sources during COVID-19 pandemic: A cross-sectional study in Japan

BACKGROUND: individual preventive behaviors are one of the key measures needed to prevent the spread of COVID-19. This study sought to identify the factors associated with the adoption of COVID-19 preventive measures, focusing specifically on information sources. METHODS: we conducted a nationally representative cross-sectional survey of 30,053 Japanese adults in February 2021. The survey asked about socioeconomic, health-related, and psychological characteristics, attitudes toward immunization, and the use of information sources regarding COVID-19. We have constructed multivariable logistic regression to estimate the factors associated with the adoption of three preventive measures: 3Cs avoidance, hand hygiene and respiratory hygiene. RESULTS: socioeconomic variables, psychological variables, and the use of information sources are significantly associated with the adoption of preventive measures. The more information sources one uses, the more likely one is to adopt preventive measures. Trust in healthcare professionals is positively associated with adopting preventive measures. On the other hand, negative correlations between trust in social media and preventive behaviors were observed. CONCLUSIONS: encouraging access to multiple information sources, utilizing communication channels, and modifying messaging according to target groups are essential to promote COVID-19 preventive measures

Business competitiveness: building and applying the 3Cs and the Strategic Change Matrix across COVID-19

This India-wide, empirical, point-in-time, global literature-supported, quantitative study involves 232 leading management consulting firm (MCF) consultants and contracting client firm (CF) perspectives. It relationally builds a structural MCF-CF 3Cs model that links MCF competencies into MCF-CF capabilities systems and then into CF business competitiveness. The 3Cs model can be visually and numerically presented as a CF three-dimensional positioning within the strategic change matrix. Future pathways towards a new optimal strategic future CF position can then be strategic change matrix mapped. A contribution towards Management-Consulting-Theory is presented as one that likely follows and embodies the MCF-CF 3Cs model processes

RNAi Nanotechnology: A Platform for siRNA Screening and Cancer Gene Therapy

Over the past two decades, advances in RNA structural biology have improved our understanding of the structures and folding properties of naturally occurring RNAs. RNA sequences and structures participate in many specific biological functions, such as those performed by messenger RNA (mRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), micro RNA (miRNA), short-interfering RNA (siRNA), small nuclear RNA (snRNA) and many others. The noncoding RNAs, such as siRNA, do not express proteins but have been utilized in a wide range of applications, including RNA interference (RNAi) and the regulation of mRNA expression. These important biological functions have been implemented in gene therapy and for screening malignant gene targets. In spite of their therapeutic potential, naturally occurring siRNAs are limited by poor pharmacological properties which has hindered their translation into the clinic. However, recent studies have highlighted the fruitful applications of modified siRNAs, including the use of siRNA nanostructures in cancer detection and treatment. In this thesis, the prerequisite conditions for forming stable RNA hybrid assemblies are described in Chapter 2. These conditions are critically important for the generation of stable higher-order RNA nanostructures. Inspired by the widespread biological function of self-assembled RNA hybrids, linear RNA templates and two complementary strands were self-assembled in order the determine the requirements for efficient RNA hybridization into stable three-component systems (3CS). In this study the RNA sequence composition and length were found to impact hybridization and self-assembly. Moreover, buffer conditions were also evaluated in order to explore the influence of ionic strength and metal cation composition on stable RNA hybridization. The complementary RNAs were annealed in buffer and analyzed by native PAGE, thermal denaturation and CD spectroscopy. The data supported the stable 3CS self-assembly on a thirty nucleotide (30nt) RNA template and with complementary 15nt and 23 nt RNA sequences in Tris buffer. These conditions were shown to favor the self-assembly of higher-order RNA structures, such as the siRNA nanostructures in Chapter 3. The genetically encoded, self-assembled siRNA nanostructures targeting the Glucose Regulated Proteins (GRP) were developed for applications in siRNA screening of these important oncologic targets and for potentiating cancer gene therapy. In our RNAi nanotechnology approach, linear, V- and Y-shape RNA templates were synthesized by semi-automated solid phase RNA synthesis with the use of a ribouridine branchpoint synthon which was used generate the V- and Y-shape RNA templates. The RNA templates were then hybridized in Tris buffer with their complementary strands, in stoichiometric ratios which favored hybridization and self-assembly into genetically encoded spheres, triangles, squares, pentagons and hexagons of discrete sizes and shapes. The siRNA self-assembly was confirmed by native PAGE while TEM imaging validated the sizes and shapes of the siRNA nanostructures. Moreover, thermal denaturation and CD spectroscopy were used to ascertain the prerequisite siRNA hybrids for their RNAi applications. In a 24 sample siRNA screen conducted within the AN3CA endometrial cancer cells known to overexpress tumorigenic GRP78 activity, the self-assembled siRNAs targeting multiple sites of GRP78 mRNA demonstrated more potent and long-lasting anticancer activity relative to their linear controls. Extending the scope of our RNAi screening approach, the self-assembled siRNA hybrids (5 nM) that targeted GRP-75, 78 and 95 were tested within endometrial (AN3CA), cervical (HeLa) and breast (MDA-MB-231) cancer cell lines with respect to the control non-cancerous lung (MRC5) cell line. The results indicated that the non-cancerous MRC5 lung cell line which displayed normal glucose regulated chaperone levels was found to tolerate siRNA treatment and demonstrated less toxicity relative to the cancer cells that were found to be addicted to glucose regulated chaperome. Therefore, the GRP targeting siRNAs were found to elicit more potent anti-cancer activity due to an overexpression and strong dependence of GRP activity in cancer. The serum stability of the self-assembled siRNAs was also investigated relative to the linear siRNA control. The data analyzed on a denaturing PAGE indicated a quick (\u3c 4 h) degradation profile of the linear siRNA hybrid while the siRNA nanostructures were disassembled into their native RNA templates with no further degradation observed over the course of a 48 h fetal bovine serum (FBS) treatment. Thus, the RNA templates have been proposed to contribute to the prolonged (72 h) RNAi effect observed within the cancer cells. In sum, these remarkable self-assembled siRNA nanostructures may thus encompass a new class of potent siRNAs that may be useful in screening important oncogene targets while improving siRNA therapeutic efficacy and specificity in cancer

RNAi Nanotechnology: A Platform for siRNA Screening and Cancer Gene Therapy

Over the past two decades, advances in RNA structural biology have improved our understanding of the structures and folding properties of naturally occurring RNAs. RNA sequences and structures participate in many specific biological functions, such as those performed by messenger RNA (mRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), micro RNA (miRNA), short-interfering RNA (siRNA), small nuclear RNA (snRNA) and many others. The noncoding RNAs, such as siRNA, do not express proteins but have been utilized in a wide range of applications, including RNA interference (RNAi) and the regulation of mRNA expression. These important biological functions have been implemented in gene therapy and for screening malignant gene targets. In spite of their therapeutic potential, naturally occurring siRNAs are limited by poor pharmacological properties which has hindered their translation into the clinic. However, recent studies have highlighted the fruitful applications of modified siRNAs, including the use of siRNA nanostructures in cancer detection and treatment. In this thesis, the prerequisite conditions for forming stable RNA hybrid assemblies are described in Chapter 2. These conditions are critically important for the generation of stable higher-order RNA nanostructures. Inspired by the widespread biological function of self-assembled RNA hybrids, linear RNA templates and two complementary strands were self-assembled in order the determine the requirements for efficient RNA hybridization into stable three-component systems (3CS). In this study the RNA sequence composition and length were found to impact hybridization and self-assembly. Moreover, buffer conditions were also evaluated in order to explore the influence of ionic strength and metal cation composition on stable RNA hybridization. The complementary RNAs were annealed in buffer and analyzed by native PAGE, thermal denaturation and CD spectroscopy. The data supported the stable 3CS self-assembly on a thirty nucleotide (30nt) RNA template and with complementary 15nt and 23 nt RNA sequences in Tris buffer. These conditions were shown to favor the self-assembly of higher-order RNA structures, such as the siRNA nanostructures in Chapter 3. The genetically encoded, self-assembled siRNA nanostructures targeting the Glucose Regulated Proteins (GRP) were developed for applications in siRNA screening of these important oncologic targets and for potentiating cancer gene therapy. In our RNAi nanotechnology approach, linear, V- and Y-shape RNA templates were synthesized by semi-automated solid phase RNA synthesis with the use of a ribouridine branchpoint synthon which was used generate the V- and Y-shape RNA templates. The RNA templates were then hybridized in Tris buffer with their complementary strands, in stoichiometric ratios which favored hybridization and self-assembly into genetically encoded spheres, triangles, squares, pentagons and hexagons of discrete sizes and shapes. The siRNA self-assembly was confirmed by native PAGE while TEM imaging validated the sizes and shapes of the siRNA nanostructures. Moreover, thermal denaturation and CD spectroscopy were used to ascertain the prerequisite siRNA hybrids for their RNAi applications. In a 24 sample siRNA screen conducted within the AN3CA endometrial cancer cells known to overexpress tumorigenic GRP78 activity, the self-assembled siRNAs targeting multiple sites of GRP78 mRNA demonstrated more potent and long-lasting anticancer activity relative to their linear controls. Extending the scope of our RNAi screening approach, the self-assembled siRNA hybrids (5 nM) that targeted GRP-75, 78 and 95 were tested within endometrial (AN3CA), cervical (HeLa) and breast (MDA-MB-231) cancer cell lines with respect to the control non-cancerous lung (MRC5) cell line. The results indicated that the non-cancerous MRC5 lung cell line which displayed normal glucose regulated chaperone levels was found to tolerate siRNA treatment and demonstrated less toxicity relative to the cancer cells that were found to be addicted to glucose regulated chaperome. Therefore, the GRP targeting siRNAs were found to elicit more potent anti-cancer activity due to an overexpression and strong dependence of GRP activity in cancer. The serum stability of the self-assembled siRNAs was also investigated relative to the linear siRNA control. The data analyzed on a denaturing PAGE indicated a quick (\u3c 4 h) degradation profile of the linear siRNA hybrid while the siRNA nanostructures were disassembled into their native RNA templates with no further degradation observed over the course of a 48 h fetal bovine serum (FBS) treatment. Thus, the RNA templates have been proposed to contribute to the prolonged (72 h) RNAi effect observed within the cancer cells. In sum, these remarkable self-assembled siRNA nanostructures may thus encompass a new class of potent siRNAs that may be useful in screening important oncogene targets while improving siRNA therapeutic efficacy and specificity in cancer

Effects of desformylflustrabromine on compulsive-like and social behaviors in mouse models of OCD and autism

Thesis (M.S.) University of Alaska Fairbanks, 2023Obsessive-Compulsive Disorder (OCD) and autism spectrum disorders (ASD) share a number of similar deficits, including altered cholinergic activity in the central nervous system (CNS), manifestation of repetitive, restricted, and compulsive-like patterns of behavior, and frequent co-morbid presentation. Desformylflustrabromine is a novel positive allosteric modulator of α4β2 nicotinic acetylcholine receptors, which has recently been shown to be able to reduce compulsive-like behavior in a non-induced mouse model of OCD. In this study, I attempted to replicate the compulsive-like behavior-reducing effects of desformylflustrabromine in the mouse model of OCD, as well as expose the model to a novel assessment; social behavior, as measured by the 3-Chamber Social test. Furthermore, I altered the model with prenatal exposure to valproic acid to induce an ASD-like construct. This ASD animal model has been used extensively to study the social deficits and neurochemistry of ASD, although limited data exists on co-morbid models. By exposing the OCD model to valproic acid, I attempted to establish two additional models; a comorbid OCD / ASD model in the compulsive-like high-activity (HA) strain and an ASD model in the non-compulsive-like low-activity (LA) strain. All mouse models and controls participated in a battery of behavioral tests quantifying compulsive-, anxiety-, and depression-like behaviors, as well as the 3-chamber social test to quantify social preference behaviors. The ASD model was not strongly established, and the desformylflustrabromine appeared to be inactive in all strains and models. I assessed potential reasons for the failure to establish a robust ASD construct in the OCD mouse model, and the failure of desformylflustrabromine to have any significant effect, in contrast with previous research using the drug in the OCD mouse model