Silicone Applicator Cleaning Improvement

Silicone is curing inside the applicator tips due to process not being standardized. Due to silicone curing inside the tips, the employees are having to repeat the cleaning process resulting in added labor expenses

Systematic comparison of nonviral gene delivery strategies for efficient co-expression of two transgenes in human mesenchymal stem cells

Background Human mesenchymal stem cells (hMSCs) are being researched for cell-based therapies due to a host of unique properties, however, genetic modification of hMSCs, accomplished through nonviral gene delivery, could greatly advance their therapeutic potential. Furthermore, expression of multiple transgenes in hMSCs could greatly advance their clinical significance for treatment of multifaceted diseases, as individual transgenes could be expressed that target separate pathogenic drivers of complex diseases. Expressing multiple transgenes can be accomplished by delivering multiple DNA vectors encoding for each transgene, or by delivering a single poly-cistronic vector that encodes for each transgene and accomplishes expression through either use of multiple promoters, an internal ribosome entry site (IRES), or a 2A peptide sequence. These different transgene expression strategies have been used to express multiple transgenes in various mammalian cells, however, they have not been fully evaluated in difficult-to-transfect primary cells, like hMSCs. This study systematically compared four transgene expression and delivery strategies for expression of two reporter transgenes in four donors of hMSCs from two tissue sources using lipid- and polymer-mediate transfection, as follows: (i) delivery of separate DNA vectors in separate nanoparticles; (ii) delivery of separate DNA vectors combined in the same nanoparticle; (iii) delivery of a bi-cistronic DNA vector with an IRES sequence via nanoparticles; and (iv) delivery of a bi-cistronic DNA vector with a dual 2A peptide sequence via nanoparticles. Results Our results indicate that expression of two transgenes in hMSCs, independent of expression or delivery strategy, is inefficient compared to expressing a single transgene. However, delivery of separate DNA vectors complexed in the same nanoparticle, or delivery of a bi-cistronic DNA vector with a dual 2A peptide sequence, significantly increased the number of hMSCs expressing both transgenes compared to other conditions tested. Conclusion Separate DNA vectors delivered in the same nanoparticle and bi-cistronic DNA vectors with dual 2A peptide sequences are highly efficient at simultaneously expressing two transgenes in multiple donors of hMSCs from different tissue sources. The data presented in this work can guide the development of hMSC transfection systems for delivery of multiple transgenes, with the goal of producing clinically relevant, genetically modified hMSCs

Glucocorticoid Priming of Nonviral Gene Delivery to hMSCs Increases Transfection by Reducing Induced Stresses

Human mesenchymal stem cells (hMSCs) are under study for cell and gene therapeutics because of their immunomodulatory and regenerative properties. Safe and efficient gene delivery could increase hMSC clinical potential by enabling expression of transgenes for control over factor production, behavior, and differentiation. Viral delivery is efficient but suffers from safety issues, while nonviral methods are safe but highly inefficient, especially in hMSCs. We previously demonstrated that priming cells with glucocorticoids (Gcs) before delivery of DNA complexes significantly increases hMSC transfection, which correlates with a rescue of transfection-induced metabolic and protein synthesis decline, and apoptosis. In this work, we show that transgene expression enhancement is mediated by transcriptional activation of endogenous hMSC genes by the cytosolic glucocorticoid receptor (cGR) and that transfection enhancement can be potentiated with a GR transcription-activation synergist. We demonstrate that the Gc-activated cGR modulates endogenous hMSC gene expression to ameliorate transfection-induced endoplasmic reticulum (ER) and oxidative stresses, apoptosis, and inflammatory responses to prevent hMSC metabolic and protein synthesis decline, resulting in enhanced transgene expression after nonviral gene delivery to hMSCs. These results provide insights important for rational design of more efficient nonviral gene delivery and priming techniques that could be utilized for clinical hMSC applications

Silicone Applicator Cleaning Improvement

Cardinal Glass is a residential glass manufacturer for doors and windows. Cardinal glass was first founded in 1962 by a man named M.L. Gordon. Today, they employ 6,000 employees across 37 manufacturing plants throughout the United States. The Greenfield, Iowa, plant distributes to local large window manufacturers. Cardinal Glass is separated into five divisions. They are as followed:FG- Float & High Volume Tempered Glass; CT- Custom Tempered Glass; LG- Laminated Glass; CG- coated glass and optical mirrors; and IG- Insulating Glass. The Client is experiencing problems with silicone curing in applicator tips due to lack of a better process. The silicone cured tips are causing re-work that is costing the company thousands of dollars in labor

The Student Movement Volume 108 Issue 1: \u2723 and me: Welcome to the AU Family!

HUMANS Babbling at the Crayon Box, Anneliese Tessalee Dorm Sweet Dorm, Savannah Tyler Surviving Freshman Year 101, Colin Cha ARTS & ENTERTAINMENT AU\u27s Reception of Barbie , Amelia Stefanescu Hey, How Was Your Summer? , Nailea Soto Sewing as an Art Form: My Experience as a First-Time Formal Dressmaker, Daena Holbrook Shadow & Bone: Reentering the Grishaverse, Madison Vath NEWS Another Generation, Another Convocation, Melissa Moore Canada\u27s Fiery Struggle: The Ongoing Battle Against Wildfires, Brendan Oh Labor Day, the Writers\u27 Strikes, and Fairness, Nathaniel Miller IDEAS Antibiotic Resistance, Sumin Lee Chapel Credits: Fair or Unfair?, Corinna Bevier From Flowers to Fires: Does Climate Change Rhetoric Need to Change?, Bella Hamann Suicide Prevention Month and the Power of Support, Reagan Westerman PULSE All That and Then Summer, Lexie Dunham Food Near AU, Alyssa Caruthers Mirror, Mirror on the Wall, is There a Fairest of Them All?, Anna Rybachek Social Media Fasts, Rodney Bell II LAST WORD You Are a God Who Sees Me, Chris Ngugihttps://digitalcommons.andrews.edu/sm-108/1000/thumbnail.jp

AD51B in Familial Breast Cancer

Common variation on 14q24.1, close to RAD51B, has been associated with breast cancer: rs999737 and rs2588809 with the risk of female breast cancer and rs1314913 with the risk of male breast cancer. The aim of this study was to investigate the role of RAD51B variants in breast cancer predisposition, particularly in the context of familial breast cancer in Finland. We sequenced the coding region of RAD51B in 168 Finnish breast cancer patients from the Helsinki region for identification of possible recurrent founder mutations. In addition, we studied the known rs999737, rs2588809, and rs1314913 SNPs and RAD51B haplotypes in 44,791 breast cancer cases and 43,583 controls from 40 studies participating in the Breast Cancer Association Consortium (BCAC) that were genotyped on a custom chip (iCOGS). We identified one putatively pathogenic missense mutation c.541C>T among the Finnish cancer patients and subsequently genotyped the mutation in additional breast cancer cases (n = 5259) and population controls (n = 3586) from Finland and Belarus. No significant association with breast cancer risk was seen in the meta-analysis of the Finnish datasets or in the large BCAC dataset. The association with previously identified risk variants rs999737, rs2588809, and rs1314913 was replicated among all breast cancer cases and also among familial cases in the BCAC dataset. The most significant association was observed for the haplotype carrying the risk-alleles of all the three SNPs both among all cases (odds ratio (OR): 1.15, 95% confidence interval (CI): 1.11–1.19, P = 8.88 x 10−16) and among familial cases (OR: 1.24, 95% CI: 1.16–1.32, P = 6.19 x 10−11), compared to the haplotype with the respective protective alleles. Our results suggest that loss-of-function mutations in RAD51B are rare, but common variation at the RAD51B region is significantly associated with familial breast cancer risk

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Reality or Illusion? The Efficacy of Nonmarket Strategy in Institutional Risk Reduction

Nonmarket strategy researchers have postulated that political and social strategies reduce the exposure of firms to risk, but those arguments have received little empirical attention. In this paper, we integrate social capital and institutional theories to examine the efficacy of managerial political ties (MPT) and corporate social responsibility (CSR) in institutional risk reduction. Using survey data from 179 firms in Ghana, we find that whereas CSR reduces institutional risk exposure, MPT do not. We also find that the effect of MPT on risk exposure is moderated by public affairs functions. Contrary to extant literature, we do not find evidence of complementarity between MPT and CSR. Altogether, the findings do not only show that the proposed efficacy of MPT in risk reduction is illusive, but they also signal the need for scrutinizing the harmony between nonmarket political and social strategie