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Reducing drug development timelines is an industry-wide goal to bring medicines to patients in need more quickly. This was exemplified in the COVID-19 pandemic where reducing development timelines had a direct impact on the number of lives lost to the disease. The use of drug substance produced using cell pools, as opposed to clones, has the potential to shorten development timelines. Toward this goal, we have developed a novel technology, GPEx® Lightning, that allows for rapid, reproducible, targeted recombination of transgenes into more than 200 Dock sites in the CHO genome. This allows for rapid production of high expressing stable cell pools and clones that reach titers of 4 to 12 g/L in generic fed-batch production. These pools and clones are highly stable in both titer and glycosylation, showing strong similarity in glycosylation profiles. This article is protected by copyright. All rights reserved.

Between the sixteenth and nineteenth century, British agriculture underwent a ‘revolutionary’ transformation. Yet despite over a century of research and the recognised centrality of agricultural developments to industrialisation and population growth, the character or chronology of any ‘revolution’ during this period remains contentious. Enquiry has been hampered by the fragmented and locally specific nature of historic accounts and the broad dating of early-modern zooarchaeological assemblages. To address this, we conducted stable isotope analysis on 658 legal documents written on sheepskin parchment; a unique biological resource that records the day, month and year of use (AD 1499 to 1969). We find these provide a high temporal resolution analysis of changing agricultural practices and episodes of disease. Most significantly, they suggest that if an ‘Agricultural Revolution’ occurred in livestock management, it did so from the mid-nineteenth century, in the aftermath of the Napoleonic Wars.

At ambient temperatures, neutral pH and ultralow concentrations (low nM), the bis(sulfonamido)bis(amido) oxidation catalyst [Fe{4-NO

The ability to control translation of endogenous or exogenous RNAs in eukaryotic cells would facilitate a variety of biotechnological applications. Current strategies are limited by low fold changes in transgene output and the size of trigger RNAs (trRNAs). Here we introduce eukaryotic toehold switches (eToeholds) as modular riboregulators. eToeholds contain internal ribosome entry site sequences and form inhibitory loops in the absence of a specific trRNA. When the trRNA is present, eToeholds anneal to it, disrupting the inhibitory loops and allowing translation. Through optimization of RNA annealing, we achieved up to 16-fold induction of transgene expression in mammalian cells. We demonstrate that eToeholds can discriminate among viral infection status, presence or absence of gene expression and cell types based on the presence of exogenous or endogenous RNA transcripts.

To examine the relationship between central lens thickness and central corneal edema during short-term closed eye scleral lens wear.Nine participants (mean age 30 years) with normal corneas wore scleral lenses (Dk 141) under closed eye conditions on separate days with nominal center thicknesses of 150, 300, 600, and 1,200 μm. Epithelial, stromal, and total corneal edema were measured using high-resolution optical coherence tomography immediately after lens application and after 90 min of wear, before lens removal. Data were corrected for variations in initial fluid reservoir thickness and compared with predictions from theoretical modeling of overnight scleral lens wear.Scleral lens-induced central corneal edema was primarily stromal in nature. The mean±standard error of corrected total corneal edema was 4.31%±0.32%, 4.55%±0.42%, 4.92%±0.50%, and 4.83%±0.22% for the 150-, 300-, 600-, and 1,200-μm lenses, respectively. No significant differences in the corrected total corneal edema were observed across all thickness groups (P=0.20). Theoretical modeling of overnight scleral lens wear seemed to overestimate the relative increase in central corneal edema as a function of decreasing lens Dk/t for values lower than 25.The magnitude of scleral lens-induced central corneal edema during short-term closed eye lens wear did not vary significantly with increasing central lens thickness. Theoretical modeling of overnight closed eye scleral lens wear seems to overestimate the effect of increasing lens thickness.

Aims: Transition Care Programs support older adults to return home post hospital admission. This study explored the experiences of older adults who identified therapy goals relating to out-of-home occupations. Methods: In this convergent mixed methods study, we assessed 10 older adults using the Meaningful Activity Participation Assessment and the Life-space Questionnaire, followed by in-depth interviews to explore out-of-home occupational engagement. Results: Participants described continued engagement in out-of-home occupations, albeit adapted due to their changing health. They reported experiencing an ongoing health and occupational journey which required them to actively consider barriers and enablers to their occupational engagement. These adults were skilled in analyzing their own performance and engagement in out-of-home occupations. Conclusion: Older adults make progressive adaptations to maintain their occupational engagement. They are experts in analyzing enablers and barriers for out-of-home occupations and should be proactively invited to share their expertise during therapy.

The steadfast advance of the synthetic biology field has enabled scientists to use genetically engineered cells, instead of small molecules or biologics, as the basis for the development of novel therapeutics. Cells endowed with synthetic gene circuits can control the localization, timing and dosage of therapeutic activities in response to specific disease biomarkers and thus represent a powerful new weapon in the fight against disease. Here, we conceptualize how synthetic biology approaches can be applied to programme living cells with therapeutic functions and discuss the advantages that they offer over conventional therapies in terms of flexibility, specificity and predictability, as well as challenges for their development. We present notable advances in the creation of engineered cells that harbour synthetic gene circuits capable of biological sensing and computation of signals derived from intracellular or extracellular biomarkers. We categorize and describe these developments based on the cell scaffold (human or microbial) and the site at which the engineered cell exerts its therapeutic function within its human host. The design of cell-based therapeutics with synthetic biology is a rapidly growing strategy in medicine that holds great promise for the development of effective treatments for a wide variety of human diseases.

Data security plays an increasingly important role in modern telecommunications. The advent of quantum computational processors presents a significant threat to today's widely employed public key encryption algorithms, necessitating the adoption of new approaches to data encryption. Whilst quantum key distribution guarantees unconditional security for cryptographic key exchange in optical communication networks, the data rate is slow (Mbit/s), especially when compared to conventional optical communication. Here we present a highly secure encryption approach in which the encryption key, generated by quantum key distribution at a rate of up to 2.9 Mbit/s, was used to seed physical layer encryption performed using time domain spectral phase encoding (TDSPE). This allowed us to demonstrate encrypted 40 Gbit/s quadrature phase shift keyed data communications over 52.3 km of installed optical fiber, which cannot be eavesdropped using brute force computational attacks. Any attempt to eavesdrop the encrypted signal in the physical layer is highly time-sensitive - the phase states must be measured and decrypted prior to optical signal attenuation, which means that the attack procedure typically needs to be completed within a few milliseconds. This work represents the first example of quantum-enhanced physical layer encryption at realistic optical data rates that is fully secure from brute force computational attacks and the first demonstration of TDSPE using continuous-wave laser source and quadrature phase shift key modulation.

Reducing drug development timelines is an industry-wide goal to bring medicines to patients in need more quickly. This was exemplified in the COVID-19 pandemic where reducing development timelines had a direct impact on the number of lives lost to the disease. The use of drug substance produced using cell pools, as opposed to clones, has the potential to shorten development timelines. Toward this goal, we have developed a novel technology, GPEx® Lightning, that allows for rapid, reproducible, targeted recombination of transgenes into more than 200 Dock sites in the CHO genome. This allows for rapid production of high expressing stable cell pools and clones that reach titers of 4 to 12 g/L in generic fed-batch production. These pools and clones are highly stable in both titer and glycosylation, showing strong similarity in glycosylation profiles.

Aim: The “2022 ACC/AHA Guideline for the Diagnosis and Management of Aortic Disease” provides recommendations to guide clinicians in the diagnosis, genetic evaluation and family screening, medical therapy, endovascular and surgical treatment, and long-term surveillance of patients with aortic disease across its multiple clinical presentation subsets (ie, asymptomatic, stable symptomatic, and acute aortic syndromes). Methods: A comprehensive literature search was conducted from January 2021 to April 2021, encompassing studies, reviews, and other evidence conducted on human subjects that were published in English from PubMed, EMBASE, the Cochrane Library, CINHL Complete, and other selected databases relevant to this guideline. Additional relevant studies, published through June 2022 during the guideline writing process, were also considered by the writing committee, where appropriate. Structure: Recommendations from previously published AHA/ACC guidelines on thoracic aortic disease, peripheral artery disease, and bicuspid aortic valve disease have been updated with new evidence to guide clinicians. In addition, new recommendations addressing comprehensive care for patients with aortic disease have been developed. There is added emphasis on the role of shared decision making, especially in the management of patients with aortic disease both before and during pregnancy. The is also an increased emphasis on the importance of institutional interventional volume and multidisciplinary aortic team expertise in the care of patients with aortic disease.

Separation and isolation of suspended submicron particles is fundamental to a wide range of applications, including desalination, chemical processing, and medical diagnostics. Ion concentration polarization (ICP), an electrokinetic phenomenon in micro-nano interfaces, has gained attention due to its unique ability to manipulate molecules or particles in suspension and solution. Less well understood, though, is the ability of this phenomenon to generate circulatory fluid flow, and how this enables and enhances continuous particle capture. Here, we perform a comprehensive study of a low-voltage ICP, demonstrating a new electrokinetic method for extracting submicron particles via flow-enhanced particle redirection. To do so, a 2D-FEM model solves the Poisson–Nernst–Planck equation coupled with the Navier–Stokes and continuity equations. Four distinct operational modes (Allowed, Blocked, Captured, and Dodged) were recognized as a function of the particle’s charges and sizes, resulting in the capture or release from ICP-induced vortices, with the critical particle dimensions determined by appropriately tuning inlet flow rates (200–800 [µm/s]) and applied voltages (0–2.5 [V]). It is found that vortices are generated above a non-dimensional ICP-induced velocity of U*=1, which represents an equilibrium between ICP velocity and lateral flow velocity. It was also found that in the case of multi-target separation, the surface charge of the particle, rather than a particle’s size, is the primary determinant of particle trajectory. These findings contribute to a better understanding of ICP-based particle separation and isolation, as well as laying the foundations for the rational design and optimization of ICP-based sorting systems.

The molecular mechanisms regulating the development and progression of alcohol use disorder (AUD) are largely unknown. While noncoding RNAs have previously been implicated as playing key roles in AUD, long-noncoding RNA (lncRNA) remains understudied in relation to AUD. In this study, we first identified ethanol-responsive lncRNAs in the mouse hippocampus that are transcriptional network hub genes. Microarray analysis of lncRNA, miRNA, circular RNA, and protein coding gene expression in the hippocampus from chronic intermittent ethanol vapor- or air- (control) exposed mice was used to identify ethanol-responsive competing endogenous RNA (ceRNA) networks. Highly interconnected lncRNAs (genes that had the strongest overall correlation to all other dysregulated genes identified) were ranked. The top four lncRNAs were novel, previously uncharacterized genes named Gm42575, 4930413E15Rik, Gm15767, and Gm33447, hereafter referred to as Pitt1, Pitt2, Pitt3, and Pitt4, respectively. We subsequently tested the hypothesis that CRISPR/Cas9 mutagenesis of the putative promoter and first exon of these lncRNAs in C57BL/6J mice would alter ethanol drinking behavior. The Drinking in the Dark (DID) assay was used to examine binge-like drinking behavior, and the Every-Other-Day Two-Bottle Choice (EOD-2BC) assay was used to examine intermittent ethanol consumption and preference. No significant differences between control and mutant mice were observed in the DID assay. Female-specific reductions in ethanol consumption were observed in the EOD-2BC assay for Pitt1, Pitt3, and Pitt4 mutant mice compared to controls. Male-specific alterations in ethanol preference were observed for Pitt1 and Pitt2. Female-specific increases in ethanol preference were observed for Pitt3 and Pitt4. Total fluid consumption was reduced in Pitt1 and Pitt2 mutants at 15% v/v ethanol and in Pitt3 and Pitt4 at 20% v/v ethanol in females only. We conclude that all lncRNAs targeted altered ethanol drinking behavior, and that lncRNAs Pitt1, Pitt3, and Pitt4 influenced ethanol consumption in a sex-specific manner. Further research is necessary to elucidate the biological mechanisms for these effects. These findings add to the literature implicating noncoding RNAs in AUD and suggest lncRNAs also play an important regulatory role in the disease.