Engineering the sequestration of carbon dioxide using microalgae

With greenhouse gas emissions (of which CO2 is the major component) being a major environmental concern, mitigation of those emissions is becoming increasingly imperative. The ability to use a fast growing, photosynthetic organism like microalgae that can survive primarily on nutrients such as sunlight and air (with increased CO2 levels) makes it a desirable agent for CO2 sequestration. The primary goal of this project is the engineering of the sequestration of CO2 using the cultivation of the microalgae species Chlorella vulgaris. Secondary goals of the project are the exploration and development of valuable by-products of the cultivation and the determination of whether utilizing microalgae to capture CO2 could be integrated economically into an industrial facility. The batch growth kinetics of the photosynthetic algal species C. vulgaris were investigated using a well-mixed stirred bioreactor. The growth rate was found to increase as the dissolved CO2 increased to 150 mg/L (10% CO2 by volume in the gas), but fell dramatically at higher concentrations. Increasing the radiant flux also increased growth rate. With a radiant flux of 32.3 mW falling directly on the 500 mL culture media, the growth rate reached up to 3.6 mg of cells/L-h. Both pH variation (5.5 - 7.0) and mass transfer rate of CO2 (KLa between 6 h-1 and 17 h-1) had little effect on growth rate. The operation of continuously stirred tank bioreactors (CSTBs) at minimum cost is a major concern for operators. In this work, a CSTB design strategy is presented where impeller stirring speed and aeration rate are optimized to meet the oxygen demand of growing cells, simultaneously minimizing the capital and operating cost. The effect of microbial species, ions in the culture medium, impeller style, as well as changing CSTB size and biomass input density on the optimum operating conditions, is examined. A study of the effects of various parameters on the CSTB design is shown. Using the kinetic data collected in the batch growth study, a novel external loop airlift photobioreactor (ELAPB) was designed and tested. A model was developed for C. vulgaris growth in the ELAPB that incorporated growth behaviour, light attenuation, mass transfer, and fluid dynamics. The model predicts biomass accumulation, light penetration, and transient CO2 concentrations, and compares predictions to experimental data for radiant fluxes of 0.075 – 1.15 W/m2 and 0 – 20% CO2 enrichment of feed air, with a 10% average error. The effect of radiant flux and CO2 concentration is presented with discussion of radial and vertical profiles along the column. For a fed-batch culture at a biomass density of 170 mg/L, the penetration of the radiant flux was found to decrease by 50% within the first 1 cm, and 75% at 2 cm. Theoretical optimum growth conditions are determined to be 0.30 W/m2 and 6% CO2 enrichment of inlet feed air. The algal culture was observed to be a workable electron acceptor in a cathodic half cell. A net potential difference of 70 mV was achieved between the growing C. vulgaris culture acting as a cathode and a 0.02 M potassium ferrocyanide anodic half cell. Surge current and power levels of 1.0 µA/mg of cell dry weight and 2.7 mW/m2 of cathode surface area were measured between these two half cells. The recently developed photosynthetic cathode was also coupled to a fermentative anode to produce a completely microbial fuel cell. Loading effects and the effect of changing culture conditions on fuel cell operation are reported. The maximum power output measured was 0.95 mW/ m2 at 90 V and 5000 ohms. A significant increase in this output is achieved with the addition of supplemental glucose to the anodic half cell and the enrichment of the feed air bubbled into the cathodic half cell with 10% CO2. Two economic feasibility studies were performed on the integration of ELAPBs into an industrial facility. These integration studies operated the ELAPBs continuously as biocathodes in coupled microbial fuel cells (MFCs) that capture CO2 from an existing 130 million L/yr bioethanol plant, while generating electrical power and yielding oil for biodiesel to provide operational revenue to offset costs. The anodes for the coupled MFCs are the existing yeast batch fermentors, and the CO2 to be sequestered comes from the existing bioethanol production. Two different design schemes were evaluated, in both cases the maximum profit was achieved with the maximum number of tall columns operated in parallel. The first design evaluated a batch bioethanol facility with off-site oil processing, and the economic feasibility is demonstrated by the positive Net Present Worth achieved over the 20 year life of the plant, at a 10% rate of return on investment. The second design, for a continuous bioethanol operation, processes both oil and algae biomass on-site, but the economics of this second process are only positive (Internal Rate of Return 9.93%.) if the government provides financial assistance in the form of generous carbon credits (a speculative $100 per tonne of CO2 not yet attained) and a 25% capital equipment grant

PHOTOSYNTHETIC – FERMENTATIVE FUEL CELL USED IN A BIOFUEL PROCESSING FACILITY

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The Emergence of Universal Immune Receptor T Cell Therapy for Cancer

Chimeric antigen receptor (CAR) T cells have shown great success in the treatment of CD19+ hematological malignancies, leading to their recent approval by the FDA as a new cancer treatment modality. However, their broad use is limited since a CAR targets a single tumor associated antigen (TAA), which is not effective against tumors with heterogeneous TAA expression or emerging antigen loss variants. Further, stably engineered CAR T cells can continually and uncontrollably proliferate and activate in response to antigen, potentially causing fatal on-target off-tumor toxicity, cytokine release syndrome, or neurotoxicity without a method of control or elimination. To address these issues, our lab and others have developed various universal immune receptors (UIRs) that allow for targeting of multiple TAAs by T cells expressing a single receptor. UIRs function through the binding of an extracellular adapter domain which acts as a bridge between intracellular T cell signaling domains and a soluble tumor antigen targeting ligand (TL). The dissociation of TAA targeting and T cell signaling confers many advantages over standard CAR therapy, such as dose control of T cell effector function, the ability to simultaneously or sequentially target multiple TAAs, and control of immunologic synapse geometry. There are currently four unique UIR platform types: ADCC-mediating Fc-binding immune receptors, bispecific protein engaging immune receptors, natural binding partner immune receptors, and anti-tag CARs. These UIRs all allow for potential benefits over standard CARs, but also bring unique engineering challenges that will have to be addressed to achieve maximal efficacy and safety in the clinic. Still, UIRs present an exciting new avenue for adoptive T cell transfer therapies and could lead to their expanded use in areas which current CAR therapies have failed. Here we review the development of each UIR platform and their unique functional benefits, and detail the potential hurdles that may need to be overcome for continued clinical translation

Natural Variation in Fc Glycosylation of HIV-Specific Antibodies Impacts Antiviral Activity

While the induction of a neutralizing antibody response against HIV remains a daunting goal, data from both natural infection and vaccine-induced immune responses suggest that it may be possible to induce antibodies with enhanced Fc effector activity and improved antiviral control via vaccination. However, the specific features of naturally induced HIV-specific antibodies that allow for the potent recruitment of antiviral activity and the means by which these functions are regulated are poorly defined. Because antibody effector functions are critically dependent on antibody Fc domain glycosylation, we aimed to define the natural glycoforms associated with robust Fc-mediated antiviral activity. We demonstrate that spontaneous control of HIV and improved antiviral activity are associated with a dramatic shift in the global antibody-glycosylation profile toward agalactosylated glycoforms. HIV-specific antibodies exhibited an even greater frequency of agalactosylated, afucosylated, and asialylated glycans. These glycoforms were associated with enhanced Fc-mediated reduction of viral replication and enhanced Fc receptor binding and were consistent with transcriptional profiling of glycosyltransferases in peripheral B cells. These data suggest that B cell programs tune antibody glycosylation actively in an antigen-specific manner, potentially contributing to antiviral control during HIV infection

Identification of Estrogen Receptor Dimer Selective Ligands Reveals Growth-Inhibitory Effects on Cells That Co-Express ERα and ERβ

Estrogens play essential roles in the progression of mammary and prostatic diseases. The transcriptional effects of estrogens are transduced by two estrogen receptors, ERα and ERβ, which elicit opposing roles in regulating proliferation: ERα is proliferative while ERβ is anti-proliferative. Exogenous expression of ERβ in ERα-positive cancer cell lines inhibits cell proliferation in response to estrogen and reduces xenografted tumor growth in vivo, suggesting that ERβ might oppose ERα's proliferative effects via formation of ERα/β heterodimers. Despite biochemical and cellular evidence of ERα/β heterodimer formation in cells co-expressing both receptors, the biological roles of the ERα/β heterodimer remain to be elucidated. Here we report the identification of two phytoestrogens that selectively activate ERα/β heterodimers at specific concentrations using a cell-based, two-step high throughput small molecule screen for ER transcriptional activity and ER dimer selectivity. Using ERα/β heterodimer-selective ligands at defined concentrations, we demonstrate that ERα/β heterodimers are growth inhibitory in breast and prostate cells which co-express the two ER isoforms. Furthermore, using Automated Quantitative Analysis (AQUA) to examine nuclear expression of ERα and ERβ in human breast tissue microarrays, we demonstrate that ERα and ERβ are co-expressed in the same cells in breast tumors. The co-expression of ERα and ERβ in the same cells supports the possibility of ERα/β heterodimer formation at physio- and pathological conditions, further suggesting that targeting ERα/β heterodimers might be a novel therapeutic approach to the treatment of cancers which co-express ERα and ERβ

Management of infantile hemangiomas during the COVID pandemic

This article is made available for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.The COVID‐19 pandemic has caused significant shifts in patient care including a steep decline in ambulatory visits and a marked increase in the use of telemedicine. Infantile hemangiomas (IH) can require urgent evaluation and risk stratification to determine which infants need treatment and which can be managed with continued observation. For those requiring treatment, prompt initiation decreases morbidity and improves long‐term outcomes. The Hemangioma Investigator Group has created consensus recommendations for management of IH via telemedicine. FDA/EMA‐approved monitoring guidelines, clinical practice guidelines, and relevant, up‐to‐date publications regarding initiation and monitoring of beta‐blocker therapy were used to inform the recommendations. Clinical decision‐making guidelines about when telehealth is an appropriate alternative to in‐office visits, including medication initiation, dosage changes, and ongoing evaluation, are included. The importance of communication with caregivers in the context of telemedicine is discussed, and online resources for both hemangioma education and propranolol therapy are provided

Genetics of circulating inflammatory proteins identifies drivers of immune-mediated disease risk and therapeutic targets

Circulating proteins have important functions in inflammation and a broad range of diseases. To identify genetic influences on inflammation-related proteins, we conducted a genome-wide protein quantitative trait locus (pQTL) study of 91 plasma proteins measured using the Olink Target platform in 14,824 participants. We identified 180 pQTLs (59 cis, 121 trans). Integration of pQTL data with eQTL and disease genome-wide association studies provided insight into pathogenesis, implicating lymphotoxin-alpha in multiple sclerosis. Using Mendelian randomization (MR) to assess causality in disease etiology, we identified both shared and distinct effects of specific proteins across immune-mediated diseases, including directionally discordant effects of CD40 on risk of rheumatoid arthritis versus multiple sclerosis and inflammatory bowel disease. MR implicated CXCL5 in the etiology of ulcerative colitis (UC) and we show elevated gut CXCL5 transcript expression in patients with UC. These results identify targets of existing drugs and provide a powerful resource to facilitate future drug target prioritization. Here the authors identify genetic effectors of the level of inflammation-related plasma proteins and use Mendelian randomization to identify proteins that contribute to immune-mediated disease risk

Designing a Library of Lived Experience for Mental Health: integrated realist synthesis and experience-based co-design study in UK mental health services

ObjectiveLiving Library events involve people being trained as living ‘Books’, who then discuss aspects of their personal experiences in direct conversation with attendees, referred to as ‘Readers’. This study sought to generate a realist programme theory and a theory informed implementation guide for a Library of Lived Experience for Mental Health (LoLEM).DesignIntegrated realist synthesis and experience-based co-design.Setting Ten online workshops with participants based in the North of England.ParticipantsThirty-one participants with a combination of personal experience of using mental health services, caring for someone with mental health difficulties, and/or working in mental health support roles.ResultsDatabase searches identified 30 published and grey literature evidence sources which were integrated with data from 10 online co-design workshops conducted over 12 months. The analysis generated a programme theory comprising five context-mechanism-outcome (CMO) configurations. Findings highlight how establishing psychological safety is foundational to productive Living Library events (CMO1). For Readers, direct conversations humanise others’ experiences (CMO 2) and provide the opportunity to flexibly explore new ways of living (CMO 3). Through participation in a Living Library, Books may experience personal empowerment (CMO 4), while the process of self- authoring and co-editing their story (CMO 5) can contribute to personal development. This programme theory informed the co-design of an implementation guide highlighting the importance of tailoring event design and participant support to the contexts in which LoLEM events are held.ConclusionsThe LoLEM has appeal across stakeholder groups and can be applied flexibly in a range of mental health-related settings. Implementation and evaluation are required to better understand the positive and negative impacts on Books and Readers.RegistrationPROSPERO CRD42022312789Strengths and limitations of this study• This study used a novel, iterative, and creative approach to integrating theory development and intervention co-design.• A key strength of this approach was the involvement of people with lived experience expertise in mental health at every stage of co-design and theory development.• The programme theory and implementation guidance were informed by analysis of research on previous Living Libraries and detailed co-design workshops, which drew on broadprofessional and personal mental health experiences.• However, few evidence sources identified by systematic searches describe Living Libraries focused specifically on mental health

Time in a Bottle: The Evolutionary Fate of Species Discrimination in Sibling Drosophila Species

Disadvantageous hybridization favors the evolution of prezygotic isolating behaviors, generating a geographic pattern of interspecific mate discrimination where members of different species drawn from sympatric populations exhibit stronger preference for members of their own species than do individuals drawn from allopatric populations. Geographic shifts in species' boundaries can relax local selection against hybridization; under such scenarios the fate of enhanced species preference is unknown. Lineages established from populations in the region of sympatry that have been maintained as single-species laboratory cultures represent cases where allopatry has been produced experimentally. Using such cultures dating from the 1950s, we assess how Drosophila pseudoobscura and D. persimilis mate preferences respond to relaxed natural selection against hybridization. We found that the propensity to hybridize generally declines with increasing time in experimental allopatry, suggesting that maintaining enhanced preference for conspecifics may be costly. However, our data also suggest a strong role for drift in determining mating preferences once secondary allopatry has been established. Finally, we discuss the interplay between populations in establishing the presence or absence of patterns consistent with reinforcement