Search

Abstract Background The perceived risk/benefit balance of prescribed and over-the-counter (OTC) medicine, as well as complementary therapies, will significantly impact on an individual’s decision-making to use medicine. For women who are pregnant or breastfeeding, this weighing of risks and benefits becomes immensely more complex because they are considering the effect on two bodies rather than one. Indeed the balance may lie in opposite directions for the mother and baby/fetus. The aim of this paper is to generate a discussion that focuses on the complexity around risk, responsibility and decision-making of medicine use by pregnant and breastfeeding women. We will also consider the competing discourses that pregnant and breastfeeding women encounter when making decisions about medicine. Discussion Women rely not only on biomedical information and the expert knowledge of their health care professionals but on their own experiences and cultural understandings as well. When making decisions about medicines, pregnant and breastfeeding women are influenced by their families, partners and their cultural societal norms and expectations. Pregnant and breastfeeding women are influenced by a number of competing discourses. “Good” mothers should manage and avoid any risks, thereby protecting their babies from harm and put their children’s needs before their own – they should not allow toxins to enter the body. On the other hand, “responsible” women take and act on medical advice – they should take the medicine as directed by their health professional. This is the inherent conflict in medicine use for maternal bodies. Summary The increased complexity involved when one body’s actions impact the body of another – as in the pregnant and lactating body – has received little acknowledgment. We consider possibilities for future research and methodologies. We argue that considering the complexity of issues for maternal bodies can improve our understanding of risk and public health education.

Abstract Background Using plant viruses to produce desirable proteins in plants allows for using non-transgenic plant hosts and if necessary, the ability to make rapid changes in the virus construct for increased or modified protein product yields. The objective of this work was the development of advanced CMV-based protein production systems to produce Acidothermus cellulolyticus endo-1, 4-β-glucanase (E1) in non-transgenic plants. Results We used two new Cucumber mosaic virus (CMV)-based vector systems for producing the green fluorescent protein (GFP) and more importantly, the Acidothermus cellulolyticus endo-1, 4-β-glucanase (E1) in non-transgenic Nicotiana benthamiana plants. These are the inducible CMVin (CMV-based inducible) and the autonomously replicating CMVar (CMV-based advanced replicating) systems. We modified a binary plasmid containing the complete CMV RNA 3 cDNA to facilitate insertion of desired sequences, and to give modifications of the subgenomic mRNA 4 leader sequence yielding several variants. Quantitative RT-PCR and immunoblot analysis showed good levels of CMV RNA and coat protein accumulation for some variants of both CMVin and CMVar. When genes for E1 or GFP were inserted in place of the CMV coat protein, both were produced in plants as shown by fluorescence (GFP) and immunoblot analysis. Enzymatic activity assays showed that active E1 was produced in plants with yields up to ~ 11 μg/g fresh weight (FW) for specific variant constructs. We also compared in vitro CMV genomic RNA reassortants, and CMV RNA 3 mutants which lacked the C’ terminal 33 amino acids of the 3A movement protein in attempts to further increase E1 yield. Taken together specific variant constructs yielded up to ~21 μg/g FW of E1 in non-transgenic plants. Conclusions Intact, active E1 was rapidly produced in non-transgenic plants by using agroinfiltration with the CMV-based systems. This reduces the time and cost compared to that required to generate transgenic plants and still gives the comparable yields of active E1. Our modifications described here, including manipulating cloning sites for foreign gene introduction, enhance the ease of use. Also, N. benthamiana, which is particularly suitable for agroinfiltration, is a very good plant for transient protein production.

Abstract Background Direct gene synthesis is becoming more popular owing to decreases in gene synthesis pricing. Compared with using natural genes, gene synthesis provides a good opportunity to optimize gene sequence for specific applications. In order to facilitate gene optimization, we have developed a stand-alone software called Visual Gene Developer. Results The software not only provides general functions for gene analysis and optimization along with an interactive user-friendly interface, but also includes unique features such as programming capability, dedicated mRNA secondary structure prediction, artificial neural network modeling, network & multi-threaded computing, and user-accessible programming modules. The software allows a user to analyze and optimize a sequence using main menu functions or specialized module windows. Alternatively, gene optimization can be initiated by designing a gene construct and configuring an optimization strategy. A user can choose several predefined or user-defined algorithms to design a complicated strategy. The software provides expandable functionality as platform software supporting module development using popular script languages such as VBScript and JScript in the software programming environment. Conclusion Visual Gene Developer is useful for both researchers who want to quickly analyze and optimize genes, and those who are interested in developing and testing new algorithms in bioinformatics. The software is available for free download at http://www.visualgenedeveloper.net.

Abstract Background Previous research has provided evidence that socioeconomic status has an impact on invasive treatments use after acute myocardial infarction. In this paper, we compare the socioeconomic inequality in the use of high-technology diagnosis and treatment after acute myocardial infarction between the US, Quebec and Belgium paying special attention to financial incentives and regulations as explanatory factors. Methods We examined hospital-discharge abstracts for all patients older than 65 who were admitted to hospitals during the 1993–1998 period in the US, Quebec and Belgium with a primary diagnosis of acute myocardial infarction. Patients' income data were imputed from the median incomes of their residential area. For each country, we compared the risk-adjusted probability of undergoing each procedure between socioeconomic categories measured by the patient's area median income. Results Our findings indicate that income-related inequality exists in the use of high-technology treatment and diagnosis techniques that is not justified by differences in patients' health characteristics. Those inequalities are largely explained, in the US and Quebec, by inequalities in distances to hospitals with on-site cardiac facilities. However, in both Belgium and the US, inequalities persist among patients admitted to hospitals with on-site cardiac facilities, rejecting the hospital location effect as the single explanation for inequalities. Meanwhile, inequality levels diverge across countries (higher in the US and in Belgium, extremely low in Quebec). Conclusion The findings support the hypothesis that income-related inequality in treatment for AMI exists and is likely to be affected by a country's system of health care.

Kaposi's sarcoma-associated herpesvirus (KSHV) is a human gammaherpesvirus associated with Kaposi's sarcoma and B cell malignancies. Like all herpesviruses, KSHV contains conserved envelope glycoproteins (gps) involved in virus binding, entry, assembly, and release from infected cells, which are also targets of the immune response. Due to the lack of a reproducible animal model of KSHV infection, the precise functions of the KSHV gps during infection in vivo are not completely known. Fortunately, a nonhuman primate (NHP) model of KSHV infection and disease has been established utilizing closely related rhesus macaque rhadinovirus (RRV) that naturally infects rhesus macaques (RM) and possesses analogous gps to KSHV. To address the roles conserved envelope gps gH and gL play during KSHV infection in vivo , we utilized the pathogenic RRV 17577 BAC to generate chimeric forms of RRV expressing KSHV gL or KSHV gH/gL, as well as an RRV mutant lacking gL expression. These viruses incorporate KSHV gH and gL into infectious virions, and although they display variable replication and differing plaque phenotypes in primary rhesus fibroblasts, they retain the ability to infect human B cells in vitro . Importantly, we also demonstrate that RRV gp chimeras can infect RM and induce the development of antibodies against KSHV. Overall, this work demonstrates that RRV gp chimeras can serve as important tools to assess the role of KSHV gH/gL in infection and disease while also providing an NHP model for testing the efficacy of KSHV gH and gL neutralizing antibodies and vaccine strategies to prevent and treat KSHV infection.IMPORTANCERhesus macaque rhadinovirus (RRV) is a rhesus macaque homolog of KSHV and serves as a model system for examining Kaposi's sarcoma-associated herpesvirus (KSHV) infection and pathogenesis in vivo . KSHV and RRV both encode conserved herpesvirus envelope glycoproteins, including gH and gL, that are important for regulating entry into host cells. In this study, we utilized the RRV BAC system to generate chimeric forms of RRV expressing KSHV gH and gL, as well as a mutant form of RRV lacking gL expression. Although these mutant and chimeric viruses can replicate in vitro , they do display growth properties different from wild-type RRV. Importantly, we demonstrate that RRV gp chimeras are capable of infecting rhesus macaques in vivo , inducing B cell hyperplasia, and promoting the development of anti-viral antibody responses that can also recognize KSHV antigens. RRV gp chimeras provide a novel system that allows for the examination of the role of KSHV gH and gL during infection in vivo .

Background: Treadmill belt perturbations have high clinical feasibility for use in perturbation-based training in older people, but their kinematic validity is unclear. This study examined the kinematic validity of treadmill belt accelerations as a surrogate for overground walkway trips during gait in older people., Methods: Thirty-eight community-dwelling older people were exposed to two unilateral belt accelerations (8 m s-2) whilst walking on a split-belt treadmill and two trips induced by a 14 cm trip-board whilst walking on a walkway with condition presentation randomised. Anteroposterior margin of stability (MoS), number of falls, and trunk and lower limb kinematics were quantified for the step prior and five recovery steps following the treadmill perturbations and the walkway trips which elicited elevating and lowering strategies., Findings: Rates of falls following the treadmill accelerations and walkway trips were 0% and 13.1%, respectively. MoS was similar during the first recovery step (P>0.05) but less negative during subsequent recovery steps following treadmill belt accelerations than walkway trips (P<0.01) regardless of recovery strategy. Excluding the first recovery step in the lowering strategy, recovery step lengths, toe clearance, maximum trunk, hip and knee angles (P<0.05) were smaller during recovery on the treadmill compared to the walkway., Interpretation: Destabilisation by treadmill belt accelerations quickly dissipated after only one recovery step but continued for multiple recovery steps following walkway trips. Smaller trunk displacement, step lengths, toe clearance and no falls on the treadmill indicate treadmill belt accelerations may not accurately simulate the biomechanical challenge of obstacle-induced trips in older people., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Jung et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Staphylococcus aureus prosthetic joint infections (PJIs) are broadly considered incurable, and clinical diagnostics that guide conservative vs. aggressive surgical treatments don't exist. Multi-omics studies in a humanized NSG-SGM3 BLT mouse model demonstrate human T cells: 1) are remarkably heterogenous in gene expression and numbers, and 2) exist as a mixed population of activated, progenitor-exhausted, and terminally-exhausted Th1/Th17 cells with increased expression of immune checkpoint proteins (LAG3, TIM-3). Importantly, these proteins are upregulated in the serum and the bone marrow of S. aureus PJI patients. A multiparametric nomogram combining high serum immune checkpoint protein levels with low proinflammatory cytokine levels (IFN-γ, IL-2, TNF-α, IL-17) revealed that TIM-3 was highly predictive of adverse disease outcomes (AUC=0.89). Hence, T cell impairment in the form of immune checkpoint expression and exhaustion could be a functional biomarker for S. aureus PJI disease outcome, and blockade of checkpoint proteins could potentially improve outcomes following surgery., Competing Interests: Conflict of interest statement GM, EMS, and MS are inventors of a patent application filed by the University of Rochester and are currently under an exclusive licensing agreement with TEx Immunetics Inc. (TEX). GM and AG are co-founders of TEX and have stock in TEX. All other authors declare that no conflict of interest exists.

Background: Iterative design and modification are used to manufacture lower limb prosthetic sockets that meet users' needs while also fulfilling safety and mobility criteria. Greater understanding of the expectations of prosthesis users regarding prosthetic fit as well as their experiences at the time of socket fitting is needed., Objectives: Describe lower limb prosthesis user perceptions and experiences of socket comfort and discomfort during their last fitting and compare this to their expectations for a perfectly fitting socket and their satisfaction with the socket they had fitted., Study Design: Mixed methods (quantitative and qualitative)., Methods: A questionnaire was designed and administered in consultation with Amputees NSW (an amputee advocacy group) and distributed through their membership database. Qualitative responses were analyzed to explore prosthesis user experiences of socket fitting, general wear, as well as socket comfort, discomfort, and satisfaction. Quantitative responses were analyzed using descriptive, nonparametric methods., Results: Thirty participants with a range of lived experiences completed the questionnaire. Reported comfort and discomfort varied across participants and indicated that expectations and experiences of fit varied and changed over time, with some participants anticipating adjustment over time would be necessary. Qualitative analysis found experiences of prosthesis socket comfort/discomfort on the day of fitting and general experiences of socket comfort related to 2 key factors-the prosthesis itself and the individual prosthesis user., Conclusions: Understanding the experiences and expectations of prosthesis users for comfort and intended activities is central to ensure success of the socket fitting approach. Investigation into fit changes over time could provide insights into follow-up requirements, user education, and funding mechanisms., (Copyright © 2024 The Authors. Published by Wolters Kluwer incorporated on behalf of The International Society for Prosthetics and Orthotics.)

Biomanufacturing enables novel sources of compounds with constant demand, such as food coloring and preservatives, as well as new compounds with peak demand, such as diagnostics and vaccines. The COVID-19 pandemic has highlighted the need for alternative sources of research materials, thrusting research on diversification of biomanufacturing platforms. Here, we show initial results exploring the walnut somatic embryogenic system expressing the recombinant receptor binding domain (RBD) and ectodomain of the spike protein (Spike) from the SARS-CoV-2 virus. Stably transformed walnut embryo lines were selected and propagated in vitro. Both recombinant proteins were detected at 3-14 µg/g dry weight of tissue culture material. Although higher yields of recombinant protein have been obtained using more conventional biomanufacturing platforms, we also report on the production of the red pigment betanin in somatic embryos, reaching yields of 650 mg/g, even higher than red beet Beta vulgaris . This first iteration shows the potential of biomanufacturing using somatic walnut embryos that can now be further optimized for different applications sourcing specialized proteins and metabolites.

Food production and pharmaceutical synthesis are posited as essential biotechnologies for facilitating human exploration beyond Earth. These technologies not only offer critical green space and food agency to astronauts but also promise to minimize mass and volume requirements through scalable, modular agriculture within closed-loop systems, offering an advantage over traditional bring-along strategies. Despite these benefits, the prevalent model for evaluating such systems exhibits significant limitations. It lacks comprehensive inventory and mass balance analyses for crop cultivation and life support, and fails to consider the complexities introduced by cultivating multiple crop varieties, which is crucial for enhancing food diversity and nutritional value. Here we expand space agriculture modeling to account for nitrogen dependence across an array of crops and demonstrate our model with experimental fitting of parameters. By adding nitrogen limitations, an extended model can account for potential interruptions in feedstock supply. Furthermore, sensitivity analysis was used to distill key consequential parameters that may be the focus of future experimental efforts., (© 2024. The Author(s).)

People often alternate between bouts of walking and running, for instance, when adults participate in recreational activities. Transitioning between activities can be challenging for prosthesis users because existing prosthetic feet are not well-suited for both tasks. Meanwhile, switching between prostheses for different tasks is often impractical. Collectively, these challenges can present barriers to physical activity participation for people with limb loss, which can negatively impact social or physical health. This work describes the development and evaluation of a passive bimodal prosthetic foot prototype with different configurations and stiffnesses for walking and running. Users rated the bimodal prosthesis higher for standing and walking compared to a running prosthesis (+2.3 for both tasks on a seven-point Likert scale). Users rated the bimodal prosthesis higher for running compared to a walking prosthesis (+1.7 and +0.5 for 2.0 and 2.5 m/s running, respectively). Changing from walking to running mode increased the device's stiffness by 23-84%, depending on the user's preference. Users could switch between bimodal prosthesis walking and running modes quickly (21.3 ± 12.0 s). Overall, the preliminary results were encouraging in terms of user satisfaction, stiffness change between modes, and mode-switching speed. These findings motivate future exploration of this bimodal prosthesis concept., (Copyright © 2024 by ASME; reuse license CC-BY 4.0.)

This review delves into the multifaceted technologies, benefits and considerations surrounding the expression of animal proteins in plants, emphasizing its potential role in advancing global nutrition, enhancing sustainability, while being mindful of the safety considerations. As the world's population continues to grow and is projected to reach 9 billion people by 2050, there is a growing need for alternative protein sources that can meet nutritional demands while minimizing environmental impact. Plant expression of animal proteins is a cutting-edge biotechnology approach that allows crops to produce proteins traditionally derived from animals, offering a sustainable and resource-efficient manner of producing these proteins that diversifies protein production and increases food security. In the United States, it will be important for there to be clear guidance in order for these technologies to reach consumers. As consumer demand for sustainable and alternative food sources rise, biotechnologies can offer economic opportunities, making this emerging technology a key player in the market landscape., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Tusé, McNulty, McDonald and Buchman.)

Walking on sloped surfaces is challenging for many lower limb prosthesis users, in part due to the limited ankle range of motion provided by typical prosthetic ankle-foot devices. Adding a toe joint could potentially benefit users by providing an additional degree of flexibility to adapt to sloped surfaces, but this remains untested. The objective of this study was to characterize the effect of a prosthesis with an articulating toe joint on the preferences and gait biomechanics of individuals with unilateral below-knee limb loss walking on slopes. Nine active prosthesis users walked on an instrumented treadmill at a +5° incline and -5° decline while wearing an experimental foot prosthesis in two configurations: a Flexible toe joint and a Locked-out toe joint. Three participants preferred the Flexible toe joint over the Locked-out toe joint for incline and decline walking. Eight of nine participants went on to participate in a biomechanical data collection. The Flexible toe joint decreased prosthesis Push-off work by 2 Joules during both incline (p = 0.008; g = -0.63) and decline (p = 0.008; g = -0.65) walking. During incline walking, prosthetic limb knee flexion at toe-off was 3° greater in the Flexible configuration compared to the Locked (p = 0.008; g = 0.42). Overall, these results indicate that adding a toe joint to a passive foot prosthesis has relatively small effects on joint kinematics and kinetics during sloped walking. This study is part of a larger body of work that also assessed the impact of a prosthetic toe joint for level and uneven terrain walking and stair ascent/descent. Collectively, toe joints do not appear to substantially or consistently alter lower limb mechanics for active unilateral below-knee prosthesis users. Our findings also demonstrate that user preference for passive prosthetic technology may be both subject-specific and task-specific. Future work could investigate the inter-individual preferences and potential benefits of a prosthetic toe joint for lower-mobility individuals., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Teater et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Patients with darker skin phototypes self-report less facial aging than their lighter-skinned counterparts. However, the association of skin phototype with the type of cosmetic procedures received, is yet to be established in a Canadian context., Objective: To compare the pattern of nonsurgical cosmetic procedures performed on people with different Fitzpatrick SPTs., Materials and Methods: Cross-sectional study of patient encounters from October 2020-April 2022. Charts and photographs were reviewed and analyzed for age, sex, SPT, and procedure type. Participants were stratified by SPT into 2 cohorts: SPT I-III and SPT IV-VI. SPTs were collapsed into groups based on definitions of "skin of color" (SPT IV-VI) in previous literature., Results: We analyzed 350 patients with mean age 43.4, of whom 320 (91%) were female and 30 (9%) were male. The SPT I-III cohort was older (mean age 45 vs 38.5 years, p < .0001) and more frequently underwent neuromodulator injection. The SPT IV-VI cohort more frequently underwent microneedling, platelet-rich plasma, or electrodessication., Conclusion: There are distinct patterns of cosmetic procedures performed. The SPT I-III cohort more commonly received procedures to manage facial aging. The SPT IV-VI cohort was younger and more commonly underwent procedures to manage hyperpigmentation., (Copyright © 2023 by the American Society for Dermatologic Surgery, Inc. Published by Wolters Kluwer Health, Inc. All rights reserved.)

Studies of human locomotion have observed asymmetries in lower-limb kinematics, especially at the more distal joints. However, it is unclear whether these asymmetries are related to functional differences between the dominant and non-dominant limb. This study aimed to determine the effect of lower-limb dominance on foot kinematics during human locomotion. Range of motion for the metatarsophalangeal joint (MPJ) and medial longitudinal arch (MLA), as well as time duration of windlass mechanism engagement, were recorded from healthy young adults (N = 12) across a range of treadmill walking and running speeds. On the group level, there were no differences in MPJ or MLA range of motion, or windlass engagement timing, between the dominant and non-dominant limb (p > 0.05). While not explained by limb dominance, between-limb differences in MPJ and MLA ranges of motion were observed for individual participants on the order of ∼2-6°, which could be clinically relevant or impact interpretation of research data., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Stair navigation is an essential and demanding form of locomotion. During stair ascent and descent, persons with lower limb loss exhibit gait characteristics which may increase their risk of falls and joint degeneration of the intact limb. To reduce deviations from typically-able-bodied gait and overloading of the intact limb for this population, one potential intervention involves modifying passive prosthetic feet by incorporating a flexible toe joint that simulates the biological metatarsophalangeal joint. In this study, we aimed to assess the user preferences and biomechanical effects of a flexible prosthetic toe joint during stair ascent and descent for persons with unilateral lower-limb loss. Nine participants with unilateral lower-limb loss were recruited (Male; Medicare Functional Classification Level: eight K4, one K3; age: 41 ± 11 years; mass: 95 ± 13 kg; height: 1.84 ± 0.05 m; mean ± SD). No significant changes in lower-limb joint mechanics were identified. Five of nine participants preferred the unmodified prosthesis with a standard carbon fiber keel for both stair ascent and descent. Varied user preferences and inconsistent changes in lower-limb joint parameters between participants highlight the importance of subject-specific analyses and individualized device prescription., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests nor personal relationships that could have inappropriately influenced this work., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Lower limb prosthesis users exhibit high rates of joint pain and disease, such as osteoarthritis, in their intact limb. Overloading of their intact limb during daily activities may be a contributing factor. Limb loading biomechanics have been extensively studied during walking, but fewer investigations into limb loading during other functional movements exist. The purpose of this study was to characterize the lower limb loading of transtibial prosthesis users during three common daily tasks: sit-to-stand, squatting, and lifting., Methods: Eight unilateral transtibial prosthesis users performed sit-to-stand (from three chair heights), squatting, and lifting a 10 kg box. Peak vertical ground reaction forces and peak knee flexion moments were computed for each limb (intact and prosthetic) to characterize limb loading and asymmetry. Ranges of motion of the intact and prosthetic ankles were also quantified., Findings: Users had greater peak ground reaction forces and knee flexion moments in their intact limb for all tasks (p < 0.02). On average, the intact limb had 36-48% greater peak ground reaction forces and 168-343% greater peak knee flexion moments compared to the prosthetic limb. The prosthetic ankle provided <10° of ankle range of motion for all tasks, less than half the range of motion provided by the intact ankle., Interpretation: Prosthesis users overloaded their intact limb during all tasks. This asymmetric loading may lead to an accumulation of damage to the intact limb joints, such as the knee, and may contribute to the development of osteoarthritis. Prosthetic design and rehabilitation interventions that promote more symmetric loading should be investigated for these tasks., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Walking is more difficult for transtibial prosthesis users, partly due to a lack of calf muscle function. Powered ankle prostheses can partially restore calf muscle function, specifically push-off power from the soleus. But one limitation of a powered ankle is that emulating the soleus does not restore the multi-articular function of the gastrocnemius. This missing function may explain elevated hip and knee muscle demands observed in individuals walking on powered ankles. These elevated demands can make walking more fatiguing and impact mobility. Adding an Artificial Gastrocnemius to a powered ankle might improve gait for prosthesis users by reducing the prosthesis-side hip and knee demands. This work investigates if an Artificial Gastrocnemius reduced prosthesis-side hip or knee demands for individuals walking with a powered ankle providing high levels of push-off. We performed two case series studies that examined the effects that a passive elastic Artificial Gastrocnemius has on joint moment-impulses when prosthesis users walked with a powered ankle. We found that hip moment-impulse was reduced during stance when walking with an Artificial Gastrocnemius for six of seven participants. The Artificial Gastrocnemius effects on knee kinetics were variable and subject-specific, but in general, it did not reduce the knee flexor or extensor demands. The Artificial Gastrocnemius should be further explored to determine if reduced hip demands improve mobility or the user's quality of life by increasing the distance they can walk, increasing walking economy, or leading to increased physical activity or community engagement., (Copyright © 2023 by ASME.)

Photoprotection is a critical health prevention strategy to reduce the deleterious effects of ultraviolet radiation (UVR) and visible light (VL). Methods of photoprotection are reviewed in this paper, with an emphasis on sunscreen. The most appropriate sunscreen formulation for personal use depends on several factors. Active sunscreen ingredients vary in their protective effect over the UVR and VL spectrum. There are dermatologic diseases that cause photosensitivity or that are aggravated by a particular action spectrum. In these situations, sunscreen suggestions can address the specific concern. Sunscreen does not represent a single entity. Appropriate personalized sunscreen selection is critical to improve compliance and clinical outcomes. Health care providers can facilitate informed product selection with awareness of evolving sunscreen formulations and counseling patients on appropriate use. This review aims to summarize different forms of photoprotection, discuss absorption of sunscreen ingredients, possible adverse effects, and disease-specific preferences for chemical, physical or oral agents that may decrease UVR and VL harmful effects., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Following transtibial amputation, a custom-built socket is the most common interface between the prosthesis and residual limb. Desire from both prosthetists and prosthesis users for improved socket fitting processes have been well documented. However, there is currently limited information available about prosthetists' experiences of how prosthetic manufacturing workflow can contribute to socket fit problems., Objectives: This study aims to determine how socket fit problems are currently detected and managed by prosthetists and to identify challenges, management strategies, and opportunities for workflow and technological innovation during prosthesis manufacture and socket fitting., Study Design: Mixed-method (quantitative and qualitative) survey., Methods: An online survey was developed and piloted in consultation with members of the Australian Orthotic Prosthetic Association. The final 25-question survey was distributed through their membership database. Mixed methods were used to analyze survey items. Qualitative items were grouped and coded under themes relating to challenges, management strategies, and opportunities. Quantitative data were analyzed using nonparametric descriptive methods., Results: Twenty-three respondents with a range of experience completed the survey. Seven of eight major Australian states/territories were represented. Primary workflow stages presenting challenges with limited strategies/solutions available to the prosthetists were roll-on liner selection, mold or cast modifications, communication with the client, and check socket fitting. Suggested solutions included improved socket-limb interface monitoring technology., Conclusions: This study provides the first insights into prosthetist-identified challenges and limitations at different stages of the socket workflow and presents a starting point for more targeted research into innovation that may assist in these processes., (Copyright © 2022 International Society for Prosthetics and Orthotics.)

Increases in global meat demands cannot be sustainably met with current methods of livestock farming, which has a substantial impact on greenhouse gas emissions, land use, water consumption, and farm animal welfare. Cultivated meat is a rapidly advancing technology that produces meat products by proliferating and differentiating animal stem cells in large bioreactors, avoiding conventional live-animal farming. While many companies are working in this area, there is a lack of existing infrastructure and experience at commercial scale, resulting in many technical bottlenecks such as scale-up of cell culture and media availability and costs. In this study, we evaluate theoretical cultivated beef production facilities with the goal of envisioning an industry with multiple facilities to produce in total 100,000,000 kg of cultured beef per year or ~0.14% of the annual global beef production. Using the computer-aided process design software, SuperPro Designer®, facilities are modeled to create a comprehensive analysis to highlight improvements that can lower the cost of such a production system and allow cultivated meat products to be competitive. Three facility scenarios are presented with different sized production reactors; ~42,000 L stirred tank bioreactor (STR) with a base case cost of goods sold (COGS) of $35/kg, ~211,000 L STR with a COGS of $25/kg, and ~262,000 L airlift reactor (ALR) with a COGS of $17/kg. This study outlines how advances in scaled up bioreactors, alternative bioreactor designs, and decreased media costs are necessary for commercialization of cultured meat products., (© 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

Purpose: The free bilamellar autograft (FBA) procedure involves harvesting a free, full-thickness section of eyelid tissue from one of the patient's healthy eyelids to reconstruct a large defect of the involved eyelid. No vascular augmentation is employed. The purpose of this study was to determine the structural and cosmetic results of this procedure., Methods: A case series was performed, looking at patients who underwent the FBA procedure for large, full-thickness eyelid defects (>50% eyelid length) between 2009 and 2020 at a single oculoplastic surgical centre. Basal cell carcinomas most frequently met criteria for the procedure. OHSN-REB waived ethics approval. All surgeries were performed by one surgeon. A single operation, with surgical steps reported in detail, was completed with follow-up documentation at ideally 1 day, 1 week, 1 month, 3 months, 6 months, and 1 year. The mean follow-up period was 28 months., Results: Thirty-one patients (17 males, 14 females, mean age 78-years-old) were included in the case series. Comorbidities included diabetes and smoking. Most patients had known basal cell carcinomas removed from the upper or lower eyelid. The mean widths of the recipient and donor sites were 18.8 and 11.5 mm, respectively. All 31 FBA surgeries resulted in structurally functional, cosmetically pleasing, and viable eyelids. Six patients developed minor graft dehiscence, 3 developed an ectropion, and 1 developed mild superficial graft necrosis secondary to frostbite, which fully recovered. Three healing phases were identified., Conclusion: This case series adds to the currently sparse data on the free bilamellar autograft procedure. The surgical technique is clearly articulated and illustrated. The FBA procedure is a simple and efficient alternative to current surgical techniques in the reconstruction of full-thickness upper and lower eyelid defects. The FBA provides functional and cosmetic success, despite the absence of an intact blood supply, with decreased operative time and faster recovery., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 McDonald, McDonald and McDonald.)

Gender biases and inequities are prevalent across many scientific fields and biomechanics is likely no exception. While progress has been made to support women in the field, especially at biomechanics society conferences, the recent COVID-19 pandemic has exacerbated professional isolation. The International Women in Biomechanics (IWB) community started in July 2020 with the mission of fostering an environment for women and other under-represented genders in biomechanics to gain year-round support, visibility, and allyship. Nearly 700 biomechanists have joined the IWB community from over 300 universities/organizations and 33 countries. Our community ranges in career stages and professions and interacts through a forum-style platform, teleconference meetings, and social media. In 2021, we conducted a survey to identify the needs, concerns, and issues faced by individuals in the IWB community. We received 144 responses from members in 16 countries. Our survey revealed three primary needs for women in biomechanics: supportive working environments, career planning support, and addressing workplace gender bias. These results, in conjunction with scientific evidence on workforce gender bias, helped us identify three key areas to meet our mission: Member Support, Community Outreach, and Empowering Allyship. Several levels of support are required in these three areas to ensure a lasting, positive, and sustainable impact on gender equity in biomechanics. We conclude by providing our perspectives on an evidence-based call to action to continue addressing gender bias and inequity at the individual, institutional, and scientific society levels. These actions can collectively enhance our allyship for women in the field of biomechanics., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Metabolic cost minimization is thought to underscore the neural control of locomotion. Yet, avoiding high muscle activation, a cause of fatigue, often outperforms energy minimization in computational predictions of human gait. Discerning the relative importance of these criteria in human walking has proved elusive, in part, because they have not been empirically decoupled. Here, we explicitly decouple whole-body metabolic cost and 'fatigue-like' muscle activation costs (estimated from electromyography) by pitting them against one another using two distinct gait tasks. When experiencing these competing costs, participants ( n = 10) chose the task that avoided overburdening muscles (fatigue avoidance) at the expense of higher metabolic power ( p < 0.05). Muscle volume-normalized activation more closely models energy use and was also minimized by the participants' decision ( p < 0.05), demonstrating that muscle activation was, at best, an inaccurate signal for metabolic energy. Energy minimization was only observed when there was no adverse effect on muscle activation costs. By decoupling whole-body metabolic and muscle activation costs, we provide among the first empirical evidence of humans embracing non-energetic optimality in favour of a clearly defined neuromuscular objective. This finding indicates that local muscle fatigue and effort may well be key factors dictating human walking behaviour and its evolution.

Microgravity-induced bone loss is a main obstacle for long term space missions as it is difficult to maintain bone mass when loading stimuli is reduced. With a typical bone mineral density loss of 1.5% per month of microgravity exposure, the chances for osteoporosis and fractures may endanger astronauts' health. Parathyroid Hormone or PTH (1-34) is an FDA approved treatment for osteoporosis, and may reverse microgravity-induced bone loss. However, PTH proteins requires refrigeration, daily subcutaneous injection, and have a short shelf-life, limiting its use in a resource-limited environment, like space. In this study, PTH was produced in an Fc-fusion form via transient expression in plants, to improve the circulatory half-life which reduces dosing frequency and to simplify purification if needed. Plant-based expression is well-suited for space medicine application given its low resource consumption and short expression timeline. The PTH-Fc accumulation profile in plant was established with a peak expression on day 5 post infiltration of 373 ± 59 mg/kg leaf fresh weight. Once the PTH-Fc was purified, the amino acid sequence and the binding affinity to its target, PTH 1 receptor (PTH1R), was determined utilizing biolayer interferometry (BLI). The binding affinity between PTH-Fc and PTH1R was 2.30 × 10 -6  M, similar to the affinity between PTH (1-34) and PTH1R (2.31 × 10 -6  M). Its function was also confirmed in a cell-based receptor stimulation assay, where PTH-Fc was able to stimulate the PTH1R producing cyclic adenosine monophosphate (cAMP) with an EC 50 of (8.54 ± 0.12) x 10 -9  M, comparable to the EC 50 from the PTH (1-34) of 1.49 × 10 -8  M. These results suggest that plant recombinant PTH-Fc exhibits a similar binding affinity and potency in a PTH1R activation assay compared to PTH. Furthermore, it can be produced rapidly at high levels with minimal resources and reagents, making it ideal for production in low resource environments such as space., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xiong, Hirano, Lane, Nandi and McDonald.)

SARS-CoV-2 Spike is a key protein that mediates viral entry into cells and elicits antibody responses. Its importance in infection, diagnostics, and vaccinations has created a large demand for purified Spike for clinical and research applications. Spike is difficult to express, prompting modifications to the protein and expression platforms to improve yields. Alternatively, the Spike receptor-binding domain (RBD) is commonly expressed with higher titers, though it has lower sensitivity in serological assays. Here, we improve transient Spike expression in Chinese hamster ovary (CHO) cells. We demonstrate that Spike titers increase significantly over the expression period, maximizing at 14 mg L -1 on day 7. In comparison, RBD titers peak at 54 mg L -1 on day 3. Next, we develop eight Spike truncations (T1-T8) in pursuit of truncation with high expression and antibody binding. The truncations T1 and T4 express at 130 and 73 mg L -1 , respectively, which are higher than our RBD titers. Purified proteins were evaluated for binding to antibodies raised against full-length Spike. T1 has similar sensitivity as Spike against a monoclonal antibody and even outperforms Spike for a polyclonal antibody. These results suggest that T1 is a promising Spike alternative for use in various applications., (© 2022 The Authors. Biotechnology Journal published by Wiley-VCH GmbH.)

Making statistical inference on quantities defining various characteristics of a temporally measured biochemical process and analyzing its variability across different experimental conditions is a core challenge in various branches of science. This problem is particularly difficult when the amount of data that can be collected is limited in terms of both the number of replicates and the number of time points per process trajectory. We propose a method for analyzing the variability of smooth functionals of the growth or production trajectories associated with such processes across different experimental conditions. Our modeling approach is based on a spline representation of the mean trajectories. We also develop a bootstrap-based inference procedure for the parameters while accounting for possible multiple comparisons. This methodology is applied to study two types of quantities-the "time to harvest" and "maximal productivity"-in the context of an experiment on the production of recombinant proteins. We complement the findings with extensive numerical experiments comparing the effectiveness of different types of bootstrap procedures for various tests of hypotheses. These numerical experiments convincingly demonstrate that the proposed method yields reliable inference on complex characteristics of the processes even in a data-limited environment where more traditional methods for statistical inference are typically not reliable.

Lower limb prosthesis users cite uneven terrain as a challenging surface to walk on. We sought to determine whether adding a Flexible toe joint to a prosthetic foot would be preferred by unilateral below-knee prosthesis users relative to a Locked (non-articulating) toe joint for walking on uneven terrain. We also quantified lower limb joint kinetics for the Locked and Flexible toe joint configurations. Five of our nine participants preferred the Flexible toe joint when walking on uneven terrain, yet from a biomechanical standpoint, the reason for this is unclear. All participants exhibited reductions in prosthetic limb net positive hip joint work when walking with a Flexible toe joint (11%; p < 0.05). For other parameters (e.g., prosthetic side knee joint moments) we observed high inter-subject variation, which adds to a growing body of literature highlighting the need for subject-specific data analyses in lower limb prosthetics research., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The virus-based immunosorbent nanoparticle is a nascent technology being developed to serve as a simple and efficacious agent in biosensing and therapeutic antibody purification. There has been particular emphasis on the use of plant virions as immunosorbent nanoparticle chassis for their diverse morphologies and accessible, high yield manufacturing via plant cultivation. To date, studies in this area have focused on proof-of-concept immunosorbent functionality in biosensing and purification contexts. Here we consolidate a previously reported pro-vector system into a single Agrobacterium tumefaciens vector to investigate and expand the utility of virus-based immunosorbent nanoparticle technology for therapeutic protein purification. We demonstrate the use of this technology for Fc-fusion protein purification, characterize key nanomaterial properties including binding capacity, stability, reusability, and particle integrity, and present an optimized processing scheme with reduced complexity and increased purity. Furthermore, we present a coupling of virus-based immunosorbent nanoparticles with magnetic particles as a strategy to overcome limitations of the immunosorbent nanoparticle sedimentation-based affinity capture methodology. We report magnetic separation results which exceed the binding capacity reported for current industry standards by an order of magnitude., Competing Interests: AS, AG, and YG were employed by Nomad Bioscience GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 McNulty, Schwartz, Delzio, Karuppanan, Jacobson, Hart, Dandekar, Giritch, Nandi, Gleba and McDonald.)

Background: Published MarginProbe (Dune Medical Devices Ltd., Israel) data reports ≥50% reduction in positive lumpectomy margins. We sought to determine whether adjunctive use of MarginProbe would provide value over intraoperative pathologic assessment alone., Methods: This is a retrospective chart review of 86 consecutive lumpectomies with MarginProbe from December 2018 to November 2019. Margins were considered positive using 'no ink on tumor' guideline for invasive cancer, and 2 mm or greater for ductal carcinoma in-situ. Significance was measured using Fisher's exact test., Results: Seventy-six patients yielded 86 lumpectomies for inclusion. Mean age was 69.8 and mean tumor size was 1.09 cm. Sixty-eight invasive cancers were assessed using adjunct MarginProbe and gross assessment, while 18 ductal carcinoma in-situ cases utilized MarginProbe only. Among all cases, gross assessment alone reduced positive margins(29.2% relative reduction, p = 0.28). Utilizing both modalities, positive margins decreased from 27.9% to 9.3% (66.7% relative reduction, p < 0.01) representing a 46.9% relative reduction versus gross assessment alone. After gross assessment and MarginProbe evaluation, additional excised volume averaged 2.9 cc., Conclusions: Synergistic use of MarginProbe and gross assessment reduces positive margins during breast conserving surgery. Surgeons can weigh its cost against it benefit with the succinct analysis we provide.

Advancements in understanding and engineering of virus-based nanomaterials (VBNs) for biomedical applications motivate a need to explore the interfaces between VBNs and other biomedically-relevant chemistries and materials. While several strategies have been used to investigate some of these interfaces with promising initial results, including VBN-containing slow-release implants and VBN-activated bioceramic bone scaffolds, there remains a need to establish VBN-immobilized three dimensional materials that exhibit improved stability and diffusion characteristics for biosensing and other analyte-capture applications. Silica sol-gel chemistries have been researched for biomedical applications over several decades and are well understood; various cellular organisms and biomolecules (e.g., bacteria, algae, enzymes) have been immobilized in silica sol-gels to improve viability, activity, and form factor (i.e., ease of use). Here we present the immobilization of an antibody-binding VBN in silica sol-gel by pore confinement. We have shown that the resulting system is sufficiently diffuse to allow antibodies to migrate in and out of the matrix. We also show that the immobilized VBN is capable of antibody binding and elution functionality under different buffer conditions for multiple use cycles. The promising results of the VBN and silica sol-gel interface indicate a general applicability for VBN-based bioseparations and biosensing applications., (© 2022. The Author(s).)

Highly detailed steered molecular dynamics simulations are performed on differently glycosylated receptor binding domains of the severe acute respiratory syndrome coronavirus-2 spike protein. The binding strength and the binding range increase with glycosylation. The interaction energy rises very quickly when pulling the proteins apart and only slowly drops at larger distances. We see a catch-slip-type behavior whereby interactions during pulling break and are taken over by new interactions forming. The dominant interaction mode is hydrogen bonds, but Lennard-Jones and electrostatic interactions are relevant as well., (Copyright © 2021 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Technoeconomic modeling and simulation is a critical step in defining a manufacturing process for evaluation of commercial viability and to focus experimental process research and development efforts. Technoeconomic analysis (TEA) is increasingly demanded alongside scientific innovation by both public and private funding agencies to maximize efficiency of resource allocation. It is particularly important for plant-based manufacturing, and other nontraditional recombinant protein production platforms, to explicitly demonstrate the manufacturing potential and to identify critical technical and economic challenges through robust technoeconomic analysis. In addition, in silico process modeling and TEA of scaled biomanufacturing facilities allows rapid evaluation of the impacts of process and economic changes on capital expenditures (CAPEX, also sometimes referred to as total capital investment), operational expenditures (OPEX, also known as total manufacturing costs or total production costs), cost of goods sold (COGS, also known as unit production costs), and profitability metrics such as net present value (NPV) and discounted cash flow rate of return (DCROR, also known as internal rate of return or IRR). These models can also be used to assess environmental, health, and safety impact of a designed biomanufacturing facility to evaluate its sustainability and environmental-friendliness. Here we describe a general method for performing technoeconomic modeling and simulation for and environmental assessment of plant-based manufacturing of recombinant proteins., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Some may think that prediction of metastasis is meaningless since metastatic breast cancer is currently incurable. We argue that effective identification of developing metastasis will enable us to design and conduct clinical trials specifically targeting those patients at high risk. The current study sought to generate the KAM score by 4 genes ( BRSK2, EYA1, SIGLEC15 , and AGTR1 ) overexpressed in primary breast cancer that developed metastasis to bone compared with matched controls without metastasis longer than 10 years. A high KAM score was prognostic of poor overall (OS), disease free survival (DFS), and disease specific survival (DSS) in the METABRIC, and OS in the GSE96058 cohorts. A high KAM score was significantly associated with clinical aggressiveness, such as high American Joint Committee Cancer (AJCC) stage, lymph node metastasis, Nottingham pathological grade, and triple negative breast cancer (TNBC). Subgroup analysis revealed that a high KAM score was associated with worse OS in ER-positive/HER2-negative breast cancer in both cohorts. A high KAM breast cancer enriched all 5 cell proliferation-related gene sets of the Hallmark collection and interferon (IFN)-γ response gene sets. Furthermore, a high KAM breast cancer was significantly infiltrated with a high fraction of not only anti-cancer but also pro-cancer immune cells and associated with high level of cytolytic activity. Finally, a high KAM breast cancer was significantly associated with lung metastasis. In conclusion, we developed KAM score using 4 gene expressions that predict lung metastasis and patient survival in breast cancer., Competing Interests: None., (AJCR Copyright © 2021.)

There are medical treatment vulnerabilities in longer-duration space missions present in the current International Space Station crew health care system with risks, arising from spaceflight-accelerated pharmaceutical degradation and resupply lag times. Bioregenerative life support systems may be a way to close this risk gap by leveraging in situ resource utilization (ISRU) to perform pharmaceutical synthesis and purification. Recent literature has begun to consider biological ISRU using microbes and plants as the basis for pharmaceutical life support technologies. However, there has not yet been a rigorous analysis of the processing and quality systems required to implement biologically produced pharmaceuticals for human medical treatment. In this work, we use the equivalent system mass (ESM) metric to evaluate pharmaceutical purification processing strategies for longer-duration space exploration missions. Monoclonal antibodies, representing a diverse therapeutic platform capable of treating multiple space-relevant disease states, were selected as the target products for this analysis. We investigate the ESM resource costs (mass, volume, power, cooling, and crew time) of an affinity-based capture step for monoclonal antibody purification as a test case within a manned Mars mission architecture. We compare six technologies (three biotic capture methods and three abiotic capture methods), optimize scheduling to minimize ESM for each technology, and perform scenario analysis to consider a range of input stream compositions and pharmaceutical demand. We also compare the base case ESM to scenarios of alternative mission configuration, equipment models, and technology reusability. Throughout the analyses, we identify key areas for development of pharmaceutical life support technology and improvement of the ESM framework for assessment of bioregenerative life support technologies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 McNulty, Berliner, Negulescu, McKee, Hart, Yates, Arkin, Nandi and McDonald.)

Transgenic rice cells (Oryza sativa) producing recombinant butyrylcholinesterase (BChE) as a prophylactic/therapeutic against organophosphate nerve agent poisoning, cocaine toxicity, and neurodegenerative diseases like Alzheimer's were immobilized in a polyethylene glycol-based hydrogel. The cells were sustained for 14 days in the semi-solid matrix, undergoing a growth phase from days 0-6, a BChE production phase in sugar-free medium from days 6-12, and a growth/recovery phase from days 12-14. Throughout this period, the cells maintained similar viability to those in suspension cultures and displayed analogous sugar consumption trends. The rice cells in the hydrogel also produced a significant amount of active BChE, comparable to the levels produced in liquid cultures. A considerable fraction of this BChE was secreted into the media, allowing for easier product separation. To the best of our knowledge, this proof-of-concept is the first report of immobilization of recombinant plant cells for continuous production of high-value heterologous proteins. This work serves as a foundation for further investigation towards plant cell bioprinting and the development of a simple, efficient, robust, modular, and potentially field-deployable bioreactor system for the manufacture of biologics., (© 2021 Wiley-VCH GmbH.)

Infectious diseases, also known as transmissible or communicable diseases, are caused by pathogens or parasites that spread in communities by direct contact with infected individuals or contaminated materials, through droplets and aerosols, or via vectors such as insects. Such diseases cause ˜17% of all human deaths and their management and control places an immense burden on healthcare systems worldwide. Traditional approaches for the prevention and control of infectious diseases include vaccination programmes, hygiene measures and drugs that suppress the pathogen, treat the disease symptoms or attenuate aggressive reactions of the host immune system. The provision of vaccines and biologic drugs such as antibodies is hampered by the high cost and limited scalability of traditional manufacturing platforms based on microbial and animal cells, particularly in developing countries where infectious diseases are prevalent and poorly controlled. Molecular farming, which uses plants for protein expression, is a promising strategy to address the drawbacks of current manufacturing platforms. In this review article, we consider the potential of molecular farming to address healthcare demands for the most prevalent and important epidemic and pandemic diseases, focussing on recent outbreaks of high-mortality coronavirus infections and diseases that disproportionately affect the developing world., (© 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

The fight against infectious diseases often focuses on epidemics and pandemics, which demand urgent resources and command attention from the health authorities and media. However, the vast majority of deaths caused by infectious diseases occur in endemic zones, particularly in developing countries, placing a disproportionate burden on underfunded health systems and often requiring international interventions. The provision of vaccines and other biologics is hampered not only by the high cost and limited scalability of traditional manufacturing platforms based on microbial and animal cells, but also by challenges caused by distribution and storage, particularly in regions without a complete cold chain. In this review article, we consider the potential of molecular farming to address the challenges of endemic and re-emerging diseases, focusing on edible plants for the development of oral drugs. Key recent developments in this field include successful clinical trials based on orally delivered dried leaves of Artemisia annua against malarial parasite strains resistant to artemisinin combination therapy, the ability to produce clinical-grade protein drugs in leaves to treat infectious diseases and the long-term storage of protein drugs in dried leaves at ambient temperatures. Recent FDA approval of the first orally delivered protein drug encapsulated in plant cells to treat peanut allergy has opened the door for the development of affordable oral drugs that can be manufactured and distributed in remote areas without cold storage infrastructure and that eliminate the need for expensive purification steps and sterile delivery by injection., (© 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Inhibitor of DNA binding ( Id ) genes comprise a family of four helix-loop-helix (HLH) transcriptional inhibitors. Our earlier studies revealed a role for ID2 within the circadian system, contributing to input, output, and core clock function through its interaction with CLOCK and BMAL1. Here, we explore the contribution of ID4 to the circadian system using a targeted disruption of the Id4 gene. Attributes of the circadian clock were assessed by monitoring the locomotor activity of Id4 -/- mice, and they revealed disturbances in its operation. Id4 -mutant mice expressed a shorter circadian period length, attenuated phase shifts in responses to continuous and discrete photic cues, and an advanced phase angle of entrainment under a 12:12 light:dark cycle and under short and long photoperiods. To understand the basis for these properties, suprachiasmatic nucleus (SCN) and retinal structures were examined. Anatomical analysis reveals a smaller Id4 -/- SCN in the width dimension, which is a finding consistent with its smaller brain. As a result of this feature, anterograde tracing in Id4 -/- mice revealed retinal afferents innovate a disproportionally larger SCN area. The Id4 -/- photic entrainment responses are unlikely to be due to an impaired function of the retinal pathways since Id4 -/- retinal anatomy and function tested by pupillometry were similar to wild-type mice. Furthermore, these circadian characteristics are opposite to those exhibited by the Id2 -/- mouse, suggesting an opposing influence of the ID4 protein within the circadian system; or, the absence of ID4 results in changes in the expression or activity of other members of the Id gene family. Expression analysis of the Id genes within the Id4 -/- SCN revealed a time-of-day specific elevated Id1 . It is plausible that the increased Id1 and/or absence of ID4 result in changes in interactions with bHLH canonical clock components or with targets upstream and/or downstream of the clock, thereby resulting in abnormal properties of the circadian clock and its entrainment.

Space missions have always assumed that the risk of spacecraft malfunction far outweighs the risk of human system failure. This assumption breaks down for longer duration exploration missions and exposes vulnerabilities in space medical systems. Space agencies can no longer reduce the majority of the human health and performance risks through crew members selection process and emergency re-supply or evacuation. No mature medical solutions exist to address this risk. With recent advances in biotechnology, there is promise for lessening this risk by augmenting a space pharmacy with a biologically-based space foundry for the on-demand manufacturing of high-value medical products. Here we review the challenges and opportunities of molecular pharming, the production of pharmaceuticals in plants, as the basis of a space medical foundry to close the risk gap in current space medical systems. Plants have long been considered to be an important life support object in space and can now also be viewed as programmable factories in space. Advances in molecular pharming-based space foundries will have widespread applications in promoting simple and accessible pharmaceutical manufacturing on Earth.

This data article is related to the research article, " M.J. McNulty, K. Kelada, D. Paul, S. Nandi, and K.A. McDonald, Introducing uncertainty quantification to techno-economic models of manufacturing field-grown plant-made products, Food Bioprod. Process. 128 (2021) 153-165." The raw and analyzed data presented are related to generation, analysis, and optimization of ultra-large-scale field-grown plant-based manufacturing of high-value recombinant protein under uncertainty. The data have been acquired using deterministic techno-economic process model simulation in SuperPro Designer integrated with stochastic Monte Carlo-based simulation in Microsoft Excel using the Crystal Ball plug-in. The purpose of the article is to make techno-economic and associated uncertainty data available to be leveraged and adapted for other research purposes., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships which have or could be perceived to have influenced the work reported in this article., (© 2021 The Authors. Published by Elsevier Inc.)

Evaluation of the functional aspects if the tumor immune microenvironment (TIME), such as the recently introduced cytolytic activity score (CYT) index have been under the spotlight in cancer research; however, clinical relevance of immune cell killing activity in breast cancer has never been analyzed in large patient cohorts. We hypothesized that CYT reflects the immune activity of TIME and can predict patient survival. A total of 7533 breast cancer patients were analyzed as both discovery and validation cohorts. We found that high CYT was associated with advanced histological grade and triple-negative breast cancer (TNBC). High CYT in tumors was significantly associated with better survival in TNBC, but unexpectedly, not in other breast cancer subtypes. High CYT TNBC included both favorable immune-related, as well as unfavorable (suppressive) inflammation-related gene sets, and characterized by high infiltration with T cells and B cells. High CYT TNBC was associated with high homologous recombination deficiency and low somatic copy number alteration score and less mutant allele tumor heterogeneity, but not with tumor mutation burden (TMB). Although CYT was not associated with pathological complete response after neoadjuvant chemotherapy, it was significantly associated with high expression of multiple immune checkpoint molecules. In conclusion, CYT of TNBC is associated with enhanced anti-cancer immunity, less intra-tumoral heterogeneity, and with better survival., Competing Interests: None., (AJCR Copyright © 2021.)