Search

Abstract Coronavirus disease 2019 (COVID-19) poses significant risks for solid organ transplant recipients, who have atypical but poorly characterized immune responses to infection. We aim to understand the host immunologic and microbial features of COVID-19 in transplant recipients by leveraging a prospective multicenter cohort of 86 transplant recipients age- and sex-matched with 172 non-transplant controls. We find that transplant recipients have higher nasal SARS-CoV-2 viral abundance and impaired viral clearance, and lower anti-spike IgG levels. In addition, transplant recipients exhibit decreased plasmablasts and transitional B cells, and increased senescent T cells. Blood and nasal transcriptional profiling demonstrate unexpected upregulation of innate immune signaling pathways and increased levels of several proinflammatory serum chemokines. Severe disease in transplant recipients, however, is characterized by a less robust induction of pro-inflammatory genes and chemokines. Together, our study reveals distinct immune features and altered viral dynamics in solid organ transplant recipients.

Abstract Mimicry of host protein structures, or ‘molecular mimicry’, is a common mechanism employed by viruses to evade the host’s immune system. Short linear amino acid (AA) molecular mimics can elicit cross-reactive antibodies and T cells from the host, but the prevalence of such mimics throughout the human virome has not been fully explored. Here we evaluate 134 human-infecting viruses and find significant usage of linear mimicry across the virome, particularly those in the Herpesviridae and Poxviridae families. Furthermore, host proteins related to cellular replication and inflammation, autosomes, the X chromosome, and thymic cells are enriched as viral mimicry targets. Finally, we find that short linear mimicry from Epstein-Barr virus (EBV) is higher in auto-antibodies found in patients with multiple sclerosis than previously appreciated. Our results thus hint that human-infecting viruses leverage mimicry in the course of their infection, and that such mimicry may contribute to autoimmunity, thereby prompting potential targets for therapies.

Cellular signaling networks are modulated by multiple protein-protein interaction domains that coordinate extracellular inputs and processes to regulate cellular processes. Several of these domains recognize short linear motifs, or SLiMs, which are often highly conserved and are closely regulated. One such domain, the Src homology 3 (SH3) domain, typically recognizes proline-rich SLiMs and is one of the most abundant SLiM-binding domains in the human proteome. These domains are often described as quite versatile, and indeed, SH3 domains can bind ligands in opposite orientations dependent on target sequence. Furthermore, recent work has identified diverse modes of binding for SH3 domains and a wide variety of sequence motifs that are recognized by various domains. Specificity is often attributed to the RT and nSrc loops near the peptide-binding cleft in this domain family, particularly for Class I binding, which is defined as RT and nSrc loop interactions with the N-terminus of the ligand. Here, we used the Src and Abl SH3 domains as a model to further investigate the role of the RT and nSrc loops in SH3 specificity. We created chimeric domains with both the RT and nSrc loop sequences swapped between these SH3 domains, and used fluorescence anisotropy assays to test how relative binding affinities were affected for Src SH3- and Abl SH3-specific ligands. We also used Alphafold–Multimer to model our SH3:peptide complexes in combination with molecular dynamics simulations. We identified a position that contributes to the nSrc loop conformation in Src SH3, the amino acid immediately following a highly conserved Trp that creates a hydrophobic pocket critical for SH3 ligand recognition. We defined this as the WX motif, where X = Trp for Src and Cys for Abl. A broad importance of this position for modulating nSrc loop conformation in SH3 domains is suggested by analyses of previously deposited SH3 structures, multiple sequence alignment of SH3 domains in the human proteome, and our biochemical and computational data of mutant Src and Abl SH3 domains. Overall, our work uses experimental approaches and structural modeling to better understand specificity determinants in SH3 domains.

Background: The underrepresentation of BIPOC youth in the fields of public health, medicine, and research may be a factor contributing to the disproportionate rates of health disparities in BIPOC communities. In 2004, the Sullivan Commission on Diversity in the Healthcare Workforce, commissioned by the White House and led by the United States Health and Human Services, recommended efforts to increase the number of minority professionals in the aforementioned fields as necessary for addressing racial and ethnic health disparities. More recently, over 240 municipalities in the United States have declared “racism a public health crisis”. This national declaration links racism directly to public health disparities, thus calling for a public health response. The Flint Public Health Youth Academy (FPHYA) provides an effective model of youth engagement steeped in Equity, Diversity, and Inclusion (EDI). FPHYA was created based on a dissertation study designed to explore the motivators for engagement of African American and other minority students into careers in public health and its six recommendations. Methods: The FPHYA Model described in this article uses a case study of the Flint Water Crisis to assess and explore effective youth engagement models for public health. This model is rooted in the Continuum of Community Engagement and Youth Empowerment Theory and explores FPHYA’s contribution of youth voice in operationalizing racism as a public health crisis.

Tree swallow nest boxes were deployed at sites proximal to two putative aqueous film forming foam (AFFF) sources in the Duluth, MN area, as well as along the St. Louis River and a reference lake for comparative purposes in 2019, 2020 and 2021. The two AFFF sites were the current Duluth Air National Guard Base (ANG) and the Lake Superior College Emergency Response Training Center. Between 13 and 40 per- and polyfluoroalkyl substances (PFAS), depending on year, were detected and quantified in tree swallow egg, nestling carcasses, and stomach contents. Assessments were made of oxidative stress and ethoxyresorufin-O-dealkylase activity in liver tissue, thyroid hormone levels in plasma and thyroid glands, DNA damage in red blood cells, and two measures of immune response (haptoglobin-like activity and immunoglobulin) in plasma of the nestlings. Additionally, other contaminants, such as polychlorinated biphenyls, legacy organochlorine pesticides, and trace elements, were assessed at sites with no previous data. Total egg PFAS concentrations at the ANG site and north of that site were 30–40 times higher than at the reference lake, while nestling PFAS concentrations were 10–15 times higher. In contrast, the St. Louis River sites had slightly, but non-statistically significant, elevated egg and nestling PFAS concentrations relative to the reference lake (2–5 times higher). One PFAS, perfluorohexane sulfonate (PFHxS), was higher, as a proportion of total PFAS, at sites with a known AFFF source compared to the reference lake, as well as compared to sites along the St. Louis River with mainly urban and industrial sources of PFAS. The ratio of total carboxylates to total sulfonates also distinguished between PFAS sources. There were few to no differences in biomarker responses among sites, and no association with PFAS exposure.

Changing climates are causing agricultural water shortages at unprecedented scales and magnitudes, especially in regions historically reliant on irrigation. Identifying and understanding systems of farming that allow continuity in agricultural operations in times of water scarcity are increasingly urgent needs. Vegetable dry farming relies on winter rains stored in soils to reduce irrigation to 0–2 events per season and has become prevalent on California’s Central Coast in recent decades. Until now, this system has been unexplored in scientific literature beyond extension publications, despite its promise as a model for low-water agriculture in arid regions. Dry farm management presents a unique challenge given that low water content restricts nutrient access in surface soils, which farmers typically target for fertility management. Managing soil nutrients at depth, as well as microorganisms that help plants access nutrients and alleviate water stress (e.g. arbuscular mycorrhizal fungi, or AMF) could be crucial to dry farm success. We engaged in a collaborative research design process with farmers managing seven commercial dry farm tomato fields to identify and answer three key management questions: 1. What are the depths at which nutrients influence harvest outcomes given low water content in surface soils?, 2. Are commercially available AMF inoculants effective at improving harvest outcomes?, and 3. How does the broader fungal community change in dry farm soils, and are those changes associated with harvest outcomes? Only soil nutrients below 60 cm depth were correlated with tomato yield and fruit quality. We identified a fungal class, Sordariomycetes, as a ‘signature’ fungal group in dry farm soils that distinguished them from irrigated management and correlated with positive fruit quality, while commercial AMF inoculation showed little benefit. These findings can inform management practices that optimize fruit yield and quality, and can guide farmers and policymakers alike in efforts to minimize agricultural water use.

Much uncertainty remains about effective messaging to boost public support for COVID-19 mitigation efforts, especially among people of color. We investigate the relationship between interview language and expressed support for COVID-19 public health protocols among Latinos: America’s largest ethnic group. Prior work establishes that interview language shapes opinions by cognitively structuring which considerations people use to express attitudes. Yet other work suggests interview language shapes opinions by activating specific cultural norms associated with a tongue. We predicted that interviewing in Spanish (versus English) would boost support for COVID-19 protocols by activating pro-social norms known to be strongly associated with that language. We uncover null support for this prediction in a pre-registered experiment on bilingual Latino adults ( N = 1645). Instead, we find that Latinos assigned to interview in Spanish report weaker support for COVID-19 protocols, regardless of which cultural norms are primed. We discuss implications for COVID-19 attitudes in linguistically diverse polities.

The transition of prebiotic chemistry to present-day chemistry lasted a very long period of time, but the current laboratory investigations of this process are mostly limited to a couple of days. Here, the authors develop a fully automated robotic prebiotic chemist designed for long-term chemical experiments exploring unconstrained multicomponent reactions, which can run autonomously and uses simple chemical inputs.

The search for life in the universe is difficult due to issues with defining signatures of living systems. Here, the authors present an approach based on the molecular assembly number and tandem mass spectrometry that allows identification of molecules produced by biological systems, and use it to identify biosignatures from a range of samples, including ones from outer space.

Summary: Background: Better understanding of the association between characteristics of patients hospitalized with coronavirus disease 2019 (COVID-19) and outcome is needed to further improve upon patient management. Methods: Immunophenotyping Assessment in a COVID-19 Cohort (IMPACC) is a prospective, observational study of 1164 patients from 20 hospitals across the United States. Disease severity was assessed using a 7-point ordinal scale based on degree of respiratory illness. Patients were prospectively surveyed for 1 year after discharge for post-acute sequalae of COVID-19 (PASC) through quarterly surveys. Demographics, comorbidities, radiographic findings, clinical laboratory values, SARS-CoV-2 PCR and serology were captured over a 28-day period. Multivariable logistic regression was performed. Findings: The median age was 59 years (interquartile range [IQR] 20); 711 (61%) were men; overall mortality was 14%, and 228 (20%) required invasive mechanical ventilation. Unsupervised clustering of ordinal score over time revealed distinct disease course trajectories. Risk factors associated with prolonged hospitalization or death by day 28 included age ≥ 65 years (odds ratio [OR], 2.01; 95% CI 1.28–3.17), Hispanic ethnicity (OR, 1.71; 95% CI 1.13–2.57), elevated baseline creatinine (OR 2.80; 95% CI 1.63– 4.80) or troponin (OR 1.89; 95% 1.03–3.47), baseline lymphopenia (OR 2.19; 95% CI 1.61–2.97), presence of infiltrate by chest imaging (OR 3.16; 95% CI 1.96–5.10), and high SARS-CoV2 viral load (OR 1.53; 95% CI 1.17–2.00). Fatal cases had the lowest ratio of SARS-CoV-2 antibody to viral load levels compared to other trajectories over time (p=0.001). 589 survivors (51%) completed at least one survey at follow-up with 305 (52%) having at least one symptom consistent with PASC, most commonly dyspnea (56% among symptomatic patients). Female sex was the only associated risk factor for PASC. Interpretation: Integration of PCR cycle threshold, and antibody values with demographics, comorbidities, and laboratory/radiographic findings identified risk factors for 28-day outcome severity, though only female sex was associated with PASC. Longitudinal clinical phenotyping offers important insights, and provides a framework for immunophenotyping for acute and long COVID-19. Funding: NIH.

Biological testing on the International Space Station (ISS) is necessary in order to monitor the microbial burden and identify risks to crew health. With support from a NASA Phase I Small Business Innovative Research contract, we have developed a compact prototype of a microgravity-compatible, automated versatile sample preparation platform (VSPP). The VSPP was built by modifying entry-level 3D printers that cost USD 200–USD 800. In addition, 3D printing was also used to prototype microgravity-compatible reagent wells and cartridges. The VSPP’s primary function would enable NASA to rapidly identify microorganisms that could affect crew safety. It has the potential to process samples from various sample matrices (swab, potable water, blood, urine, etc.), thus yielding high-quality nucleic acids for downstream molecular detection and identification in a closed-cartridge system. When fully developed and validated in microgravity environments, this highly automated system will allow labor-intensive and time-consuming processes to be carried out via a turnkey, closed system using prefilled cartridges and magnetic particle-based chemistries. This manuscript demonstrates that the VSPP can extract high-quality nucleic acids from urine (Zika viral RNA) and whole blood (human RNase P gene) in a ground-level laboratory setting using nucleic acid-binding magnetic particles. The viral RNA detection data showed that the VSPP can process contrived urine samples at clinically relevant levels (as low as 50 PFU/extraction). The extraction of human DNA from eight replicate samples showed that the DNA extraction yield is highly consistent (there was a standard deviation of 0.4 threshold cycle when the extracted and purified DNA was tested via real-time polymerase chain reaction). Additionally, the VSPP underwent 2.1 s drop tower microgravity tests to determine if its components are compatible for use in microgravity. Our findings will aid future research in adapting extraction well geometry for 1 g and low g working environments operated by the VSPP. Future microgravity testing of the VSPP in the parabolic flights and in the ISS is planned.

Summary: The characterization of the chemistry of life on earth has been facilitated by developments in analysis and sequencing of bio-oligomers using tandem mass spectrometry (MS/MS). Bio-oligomers can be identified with sequence-level resolution in analytes more complex than any synthetic mixture, enabled by well-established knowledge of fragmentation properties and extensive MS/MS databases built up over decades. However, unknown oligomer systems remain difficult to characterize, as no comparable databases exist, partly because of the vast chemical diversity and fragmentation pathways. Here, we present oligomer-soup-sequencing (OLIGOSS), a new approach to the sequencing of unknown oligomer systems. Using a novel set of backbone-agnostic abstract properties to define fragmentation, OLIGOSS is capable of sequencing any linear oligomer class amenable to MS/MS, regardless of backbone chemistry. We validated OLIGOSS by sequencing synthetic peptides, polyesters, polyimines and depsipeptide oligomers, mapped RNA methylation sites, and a ribosomally synthesized peptide, thioholgamide, directly from a cell lysate without purification.

K + channels function in macromolecular complexes with accessory subunits to regulate neuronal function. Here, the authors describe Pin1-mediated regulation of the Kv4.2 complex, which impacts reversal learning in mice, providing potential treatment for disorders characterized by cognitive inflexibility.

Growth in open-source hardware designs combined with the decreasing cost of high-quality 3D printers have supported a resurgence of in-house custom lab equipment development. Herein, we describe a low-cost (< $400), open-source CO2 incubator. The system is comprised of a Raspberry Pi computer connected to a 3D printer controller board that has controls for a CO2 sensor, solenoid valve, heater, and thermistors. CO2 is supplied through the sublimation of dry ice stored inside a thermos to create a sustained 5% CO2 supply. The unit is controlled via G-Code commands sent by the Raspberry Pi to the controller board. In addition, we built a custom software application for remote control and used the open-source Grafana dashboard for remote monitoring. Our data show that we can maintain consistent CO2 and temperature levels for over three days without manual interruption. The results from our culture plates and real-time PCR indicate that our incubator performed equally well when compared to a much more expensive commercial CO2 incubator. We have also demonstrated that the antibiotic susceptibility assay can be performed in this low-cost CO2 incubator. Our work also indicates that the system can be connected to incubator chambers of various chamber volumes.

A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD.

Bartonella bacilliformis (B. bacilliformis), Bartonella henselae (B. henselae), and Bartonella quintana (B. quintana) are bacteria known to cause verruga peruana or bacillary angiomatosis, vascular endothelial growth factor (VEGF)-dependent cutaneous lesions in humans. Given the bacteria’s association with the dermal niche and clinical suspicion of occult infection by a dermatologist, we determined if patients with melanoma had evidence of Bartonella spp. infection. Within a one-month period, eight patients previously diagnosed with melanoma volunteered to be tested for evidence of Bartonella spp. exposure/infection. Subsequently, confocal immunohistochemistry and PCR for Bartonella spp. were used to study melanoma tissues from two patients. Blood from seven of the eight patients was either seroreactive, PCR positive, or positive by both modalities for Bartonella spp. exposure. Subsequently, Bartonella organisms that co-localized with VEGFC immunoreactivity were visualized using multi-immunostaining confocal microscopy of thick skin sections from two patients. Using a co-culture model, B. henselae was observed to enter melanoma cell cytoplasm and resulted in increased vascular endothelial growth factor C (VEGFC) and interleukin 8 (IL-8) production. Findings from this small number of patients support the need for future investigations to determine the extent to which Bartonella spp. are a component of the melanoma pathobiome.

Quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay is the gold standard recommended to test for acute SARS-CoV-2 infection. However, it generally requires expensive equipment such as RNA isolation instruments and real-time PCR thermal cyclers. As a pandemic, COVID-19 has spread indiscriminately, and many low resource settings and developing countries do not have the means for fast and accurate COVID-19 detection to control the outbreak. Additionally, long assay times, in part caused by slow sample preparation steps, have created a large backlog when testing patient samples suspected of COVID-19. With many PCR-based molecular assays including an extraction step, this can take a significant amount of time and labor, especially if the extraction is performed manually. Using COVID-19 clinical specimens, we have collected evidence that the RT-qPCR assay can feasibly be performed directly on patient sample material in virus transport medium (VTM) without an RNA extraction step, while still producing sensitive test results. If RNA extraction steps can be omitted without significantly affecting clinical sensitivity, the turn-around time of COVID-19 tests, and the backlog we currently experience can be reduced drastically. Furthermore, our data suggest that rapid RT-PCR can be implemented for sensitive and specific molecular diagnosis of COVID-19 in locations where sophisticated laboratory instruments are not available. Our USD 300 set up achieved rapid RT-PCR using thin-walled PCR tubes and a water bath setup using sous vide immersion heaters, a Raspberry Pi computer, and a single servo motor that can process up to 96 samples at a time. Using COVID-19 positive clinical specimens, we demonstrated that RT-PCR assays can be performed in as little as 12 min using untreated samples, heat-inactivated samples, or extracted RNA templates with our low-cost water bath setup. These findings can help rapid COVID-19 testing to become more accessible and attainable across the globe.

The type of stretch activated receptors (SARs) in the chordotonal organs in the crab walking leg and of the muscle receptor organ (MRO) in the crayfish abdomen have not yet been classified as to their molecular or pharmacological profile. The purpose of this study is to examine the pharmacological profile of SARs in the proprioceptive neurons in the crab and crayfish models. Since many SARs share the pharmacological profile of displaying low pH or being proton sensitive (i.e. being more active) or blocked by the diuretic amiloride or ruthenium red as well as being blocked by the broad stretch activated channel blocker gadolinium (Gd3+), we used these agents to screen the receptors. Various displacement rates as well as static positions that activate the stretch activated receptors were used in examining their pharmacological profiles. Hour-long exposure to low pH decreased neural activity of the chordotonal organ of the crab more so than to amiloride or ruthenium red. The crayfish MRO did not show pH sensitivity or sensitivity to amiloride or ruthenium red. Gd3+ rapidly blocked neural activity in both the crab and crayfish. It appears these stretch activated receptors may not have a classification that is suited to the standard pharmacological profiles. The molecular makeup of the channels also awaits characterization. This could reveal a novel SAR subtype. Our neurophysiology course1 took this project on as a course-based undergraduate research experience (CURE) to address an authentic research question.

Herein, we describe the preparation, characterization and photophysical properties of neutral lanthanide complexes containing a redox-active ligand 1-(2-pyridylazo)-2-phenanthrol (papl). The complexes likely share similar structural features and bear the formulation Ln(papl)3 (Ln(III) = Gd, Dy, Tb), which is supported by electrospray ionization mass spectrometry, CHN analysis, FT-IR and UV–Vis spectroscopy. The synthesis and structural properties of a related complex, Ho(qapl)3 (where qapl = 10-(8-quinolylazo)-9-phenanthrol), is also reported. The complexes feature ligand-centered redox activity, similar to other reported transition metal complexes with papl. Variable temperature magnetic susceptibility measurements (DC and AC) suggest typical free-ion magnetism without any slow-relaxation dynamics. The photophysical properties of the ligand and complexes were investigated and the results of emission spectroscopy indicate ligand-centered processes.

Cooperation is essential for evolution of biological complexity. Recent work has shown game theoretic arguments, commonly used to model biological cooperation, can also illuminate the dynamics of chemical systems. Here we investigate the types of cooperation possible in a real RNA system based on the Azoarcus ribozyme, by constructing a taxonomy of possible cooperative groups. We construct a computational model of this system to investigate the features of the real system promoting cooperation. We find triplet interactions among genotypes are intrinsically biased towards cooperation due to the particular distribution of catalytic rate constants measured empirically in the real system. For other distributions cooperation is less favored. We discuss implications for understanding cooperation as a driver of complexification in the origin of life.

Abstract Background In humans, dynamic contrast CMR of the first pass of a bolus infusion of Gadolinium-based contrast agent has become a standard technique to identify under-perfused regions of the heart and can accurately demonstrate the severity of myocardial infarction. Despite the clinical importance of this method, it has rarely been applied in small animal models of cardiac disease. In order to identify perfusion delays in the infarcted rat heart, here we present a method in which a T1 weighted MR image has been acquired during each cardiac cycle. Methods and results In isolated perfused rat hearts, contrast agent infusion gave uniform signal enhancement throughout the myocardium. Occlusion of the left anterior descending coronary artery significantly reduced the rate of signal enhancement in anterior regions of the heart, demonstrating that the first-pass method was sensitive to perfusion deficits. In vivo measurements of myocardial morphology, function, perfusion and viability were made at 2 and 8 days after infarction. Morphology and function were further assessed using cine-MRI at 42 days. The perfusion delay was larger in rat hearts that went on to develop greater functional impairment, demonstrating that first-pass CMR can be used as an early indicator of infarct severity. First-pass CMR at 2 and 8 days following infarction better predicted outcome than cardiac ejection fraction, end diastolic volume or end systolic volume. Conclusion First-pass CMR provides a predictive measure of the severity of myocardial impairment caused by infarction in a rodent model of heart failure.

Abstract Background Conditional expression vectors have become a valuable research tool to avoid artefacts that may result from traditional gene expression studies. However, most systems require multiple plasmids that must be independently engineered into the target system, resulting in experimental delay and an increased potential for selection of a cell subpopulation that differs significantly from the parental line. We have therefore developed pHUSH, an inducible expression system that allows regulated expression of shRNA, miRNA or cDNA cassettes on a single viral vector. Results Both Pol II and Pol III promoters have been successfully combined with a second expression cassette containing a codon-optimized tetracycline repressor and selectable marker. We provide examples of how pHUSH has been successfully employed to study the function of target genes in a number of cell types within in vitro and in vivo assays, including conditional gene knockdown in a murine model of brain cancer. Conclusion We have successfully developed and employed a single vector system that enables Doxycycline regulated RNAi or transgene expression in a variety of in vitro and in vivo model systems. These studies demonstrate the broad application potential of pHUSH for conditional genetic engineering in mammalian cells.

Abstract Background Family practitioners take care of large numbers of seniors with increasingly complex mental health problems. Varying levels of input may be necessary from psychiatric consultants. This study examines patients'/family, family practitioners', and psychiatrists' perceptions of the bi-directional pathway between such primary care doctors and consultants. Methods An 18 month survey was conducted in an out-patient psychogeriatric clinic of a Montreal university-affiliated community hospital. Cognitively intact seniors referred by family practitioners for assessment completed a satisfaction and expectation survey following their visits with the psychiatric consultants. The latter completed a self-administered process of care questionnaire at the end of the visit, while family doctors responded to a similar survey by telephone after the consultants' reports had been received. Responses of the 3 groups were compared. Results 101 seniors, referred from 63 family practitioners, met the study entry criteria for assessment by 1 of 3 psychogeriatricians. Both psychiatrists and family doctors agreed that help with management was the most common reason for referral. Family physicians were accepting of care of elderly with mental health problems, but preferred that the psychiatrists assume the initial treatment; the consultants preferred direct return of the patient; and almost 1/2 of patients did not know what to expect from the consultation visit. The rates of discordance in expectations were high when each unique patient-family doctor-psychiatrist triad was examined. Conclusion Gaps in expectations exist amongst family doctors, psychiatrists, and patients/family in the shared mental health care of seniors. Goals and anticipated outcomes of psychogeriatric consultation require better definition.

Summary: Background: Increasing atmospheric concentrations of carbon dioxide (CO2) affect global nutrition via effects on agricultural productivity and nutrient content of food crops. We combined these effects with economic projections to estimate net changes in nutrient availability between 2010 and 2050. Methods: In this modelling study, we used the International Model for Policy Analysis of Agricultural Commodities and Trade to project per capita availability of protein, iron, and zinc in 2050. We used estimated changes in productivity of individual agricultural commodities to model effects on production, trade, prices, and consumption under moderate and high greenhouse gas emission scenarios. Two independent sources of data, which used different methodologies to determine the effect of increased atmospheric CO2 on different key crops, were combined with the modelled food supply results to estimate future nutrient availability. Findings: Although technological change, market responses, and the effects of CO2 fertilisation on yield are projected to increase global availability of dietary protein, iron, and zinc, these increases are moderated by negative effects of climate change affecting productivity and carbon penalties on nutrient content. The carbon nutrient penalty results in decreases in the global availability of dietary protein of 4·1%, iron of 2·8%, and zinc of 2·5% as calculated using one dataset, and decreases in global availability of dietary protein of 2·9%, iron of 3·9%, and zinc of 3·4% using the other dataset. The combined effects of projected increases in atmospheric CO2 (ie, carbon nutrient penalty, CO2 fertilisation, and climate effects on productivity) will decrease growth in the global availability of nutrients by 19·5% for protein, 14·4% for iron, and 14·6% for zinc relative to expected technology and market gains by 2050. The many countries that currently have high levels of nutrient deficiency would continue to be disproportionately affected. Interpretation: This approach is an improvement in estimating future global food security by simultaneously projecting climate change effects on crop productivity and changes in nutrient content under increased concentrations of CO2, which accounts for a much larger effect on nutrient availability than CO2 fertilisation. Regardless of the scenario used to project future consumption patterns, the net effect of increasing concentrations of atmospheric CO2 will slow progress in decreasing global nutrient deficiencies. Funding: US Environmental Protection Agency, Consultative Group on International Agricultural Research (CIGAR) Research Program on Policies, Institutions and Markets (PIM), and the CGIAR Research Program on Climate Change and Food Security (CCAFS).

Abstract: Complement-mediated thrombotic microangiopathy (CM-TMA) or hemolytic uremic syndrome, previously identified as atypical hemolytic uremic syndrome, is a TMA characterized by germ line variants or acquired antibodies to complement proteins and regulators. Building upon our prior experience with the modified Ham (mHam) assay for ex vivo diagnosis of complementopathies, we have developed an array of cell-based complement "biosensors" by selective removal of complement regulatory proteins (CD55 and CD59, CD46, or a combination thereof) in an autonomously bioluminescent HEK293 cell line. These biosensors can be used as a sensitive method for diagnosing CM-TMA and monitoring therapeutic complement blockade. Using specific complement pathway inhibitors, this model identifies immunoglobulin M (IgM)-driven classical pathway stimulus during both acute disease and in many patients during clinical remission. This provides a potential explanation for ∼50% of patients with CM-TMA who lack an alternative pathway "driving" variant and suggests at least a subset of CM-TMA is characterized by a breakdown of IgM immunologic tolerance., (© 2024 American Society of Hematology. Published by Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.)

Introduction: Passive leg raise (PLR) is a simple, dynamic maneuver that has been used to increase preload to the heart. We hypothesize that PLR may offer a new and efficient office-based tool for assessing blood pressure (BP) control in older adults., Methods: One hundred and three veterans (≥60 years old) without known cardiovascular disease and varying degrees of blood pressure control were included in this cross-sectional cohort study. Twenty-four hour ambulatory BP monitoring identified Veterans with optimal and suboptimal BP control (≥125/75 mmHg). Bioimpedance electrodes (Baxter Medical, Deerfield, Illinois, USA) and brachial BP were used to calculate hemodynamic parameter changes across PLR states [pre-PLR, active PLR (3 min), and post-PLR]. Multiple linear regression was used to assess associations between BP control status with changes in hemodynamic parameters between PLR states., Results: The 24-h ambulatory BP monitoring identified 43 (42%) older Veterans with optimal BP control (mean age of 70.5 ± 7.0 years) and 55 (54%) with suboptimal BP (mean age of 71.3 ± 8.7 years). Veterans with suboptimal BP control had significantly reduced change in total peripheral resistance (ΔTPR) (7.0 ± 156.0 vs. 127.3 ± 145.6 dynes s/cm 5 ; P  = 0.002) following PLR compared with Veterans with optimal BP control. Suboptimal BP control ( β  = -0.35, P  = 0.004) had a significant association with reduced ΔTPR, even after adjusting for demographic variables., Conclusion: Measuring PLR-induced hemodynamic changes in the office setting may represent an alternative way to identify older adults with suboptimal BP control when 24-h ambulatory BP monitoring is not available., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

The nuclear genome is spatially organized into a three-dimensional (3D) architecture by physical association of large chromosomal domains with subnuclear compartments including the nuclear lamina at the radial periphery and nuclear speckles within the nucleoplasm 1-5 . However, how spatial genome architecture regulates human brain development has been overlooked owing to technical limitations. Here, we generate high-resolution maps of genomic interactions with the lamina and speckles in cells of the neurogenic lineage isolated from midgestational human cortex, uncovering an intimate association between subnuclear genome compartmentalization, chromatin state and transcription. During cortical neurogenesis, spatial genome organization is extensively remodeled, relocating hundreds of neuronal genes from the lamina to speckles including key neurodevelopmental genes bivalent for H3K27me3 and H3K4me3. At the lamina, bivalent genes have exceptionally low expression, and relocation to speckles enhances resolution of bivalent chromatin to H3K4me3 and increases transcription >7-fold. We further demonstrate that proximity to the nuclear periphery - not the presence of H3K27me3 - is the dominant factor in maintaining the lowly expressed, poised state of bivalent genes embedded in the lamina. In addition to uncovering a critical role of subnuclear genome compartmentalization in neurogenic transcriptional regulation, our results establish a new paradigm in which knowing the spatial location of a gene is necessary to understanding its epigenomic regulation., Competing Interests: Declaration of Interests S.H.A. and D.A.L. are inventors on a provisional patent related to the methodological aspects of this study. US patent application, 63/448,001, was filed by The Regents of the University of California on February 24, 2023, entitled “Methods for Epigenetic Analysis'. Related international application, PCT/US24/16976 was filed on February 23, 2024.

Complement-mediated hemolytic uremic syndrome (CM-HUS) is a thrombotic microangiopathy characterized by germline variants or acquired antibodies to complement proteins and regulators. Building upon our prior experience with the modified Ham (mHam) assay for ex vivo diagnosis of complementopathies, we have developed an array of cell-based complement "biosensors'' by selective removal of complement regulatory proteins (CD55 and CD59, CD46, or a combination thereof) in an autonomously bioluminescent HEK293 cell line. These biosensors can be used as a sensitive method for diagnosing CM-HUS and monitoring therapeutic complement blockade. Using specific complement pathway inhibitors, this model identifies IgM-driven classical pathway stimulus during both acute disease and in many patients during clinical remission. This provides a potential explanation for ~50% of CM-HUS patients who lack an alternative pathway "driving" variant and suggests at least a subset of CM-HUS is characterized by a breakdown of IgM immunologic tolerance., Key Points: CM-HUS has a CP stimulus driven by polyreactive IgM, addressing the mystery of why 40% of CM-HUS lack complement specific variantsComplement biosensors and the bioluminescent mHam can be used to aid in diagnosis of CM-HUS and monitor complement inhibitor therapy.

Complement is a major driver of antiphospholipid syndrome (APS) and a promising therapeutic target in refractory and catastrophic APS. Complement testing in APS is largely limited to research settings, and reliable, rapid-turnaround biomarkers are needed to predict those at risk for adverse clinical outcomes and most likely to benefit from complement inhibition. We review complement biomarkers and their association with thrombosis and obstetric outcomes, including: (i) complement proteins and activation fragments in the fluid phase; (ii) assays that evaluate complement on cell membranes (e.g. in vivo cell-bound complement fragments, hemolytic assays, and ex vivo 'functional' cell-based assays, and (iii) sequencing of complement genes. Current studies highlight the inconsistencies in testing both between studies and various aPL/APS subgroups, suggesting that either cell-based testing or multiplex panels employing a combination of biomarkers simultaneously may be most clinically relevant. Standardization of complement assays is needed to ensure reproducibility and establish clinically relevant applications., Competing Interests: Declaration of Competing Interest SC has participated on advisory boards or received consulting fees from Alexion, Sanofi, UCB, Sobi, and Takeda, and has received honoraria/royalties from UpToDate.com and Dynamed.com. G.F.G. has served on advisory boards for Apellis Pharmaceuticals and Alexion Pharmaceuticals and contributed to the Merck Manual, for which she received an honorarium. G.F.G (spouse) is an employee of Pfizer and holds stock. M.A.C. has no relevant conflicts of interest to declare., (Copyright © 2023. Published by Elsevier Inc.)

Heating is a major source of air pollution. To improve air quality, a range of clean heating policies were implemented in China over the past decade. Here, we evaluated the impacts of winter heating and clean heating policies on air quality in China using a novel, observation-based causal inference approach. During 2015-2021, winter heating causally increased annual PM 2.5 , daily maximum 8-h average O 3 , and SO 2 by 4.6, 2.5, and 2.3 μg m -3 , respectively. From 2015 to 2021, the impacts of winter heating on PM 2.5 in Beijing and surrounding cities (i.e., "2 + 26" cities) decreased by 5.9 μg m -3 (41.3%), whereas that in other northern cities only decreased by 1.2 μg m -3 (12.9%). This demonstrates the effectiveness of stricter clean heating policies on PM 2.5 in "2 + 26" cities. Overall, clean heating policies caused the annual PM 2.5 in mainland China to reduce by 1.9 μg m -3 from 2015 to 2021, potentially avoiding 23,556 premature deaths in 2021.

Abstract: How can nursing academia impart concepts of spirituality, social justice, and personal and professional growth to nursing students? The authors of this qualitative study explored the narratives of 21 nursing students' reflective journals written while students were participating in a global clinical mission. A guided reflective journal grounded in Catholic social teachings and St. Francis' social justice principles was used daily during the mission to capture participants' thoughts and feelings. Data analysis revealed that students discovered principles of spirituality and social justice and expressed a deeper appreciation for being present for others, providing healthcare to all equally, and promoting the common good. Guided questions enhanced the experiential learning experience, fostering personal and professional growth., (Copyright © 2023 InterVarsity Christian Fellowship.)

Human chromosomes are pervasively transcribed, but systematic understanding of coding and lncRNA genome function in cell differentiation is lacking. Using CRISPR interference (CRISPRi) in human induced pluripotent stem cells, we performed dual genome-wide screens - assessing 18,905 protein-coding and 10,678 lncRNA loci - and identified 419 coding and 201 lncRNA genes that regulate neural induction. Integrative analyses revealed distinct properties of coding and lncRNA genome function, including a 10-fold enrichment of lncRNA genes for roles in differentiation compared to proliferation. Further, we applied Perturb-seq to obtain granular insights into neural induction phenotypes. While most coding hits stalled or aborted differentiation, lncRNA hits were enriched for the genesis of diverse cellular states, including those outside the neural lineage. In addition to providing a rich resource (danlimlab.shinyapps.io/dualgenomewide) for understanding coding and lncRNA gene function in development, these results indicate that the lncRNA genome regulates lineage commitment in a manner fundamentally distinct from coding genes., Competing Interests: DECLARATION OF INTERESTS The authors declare the following competing financial interests: The Regents of the University of California with J.S.W. and M.A.H. as inventors have filed patent applications related to CRISPRi/a screening and Perturb-seq (11,254,933). J.S.W. declares outside interest in 5AM Ventures, Amgen, Chroma Medicine, KSQ Therapeutics, Maze Therapeutics, Tenaya Therapeutics, Tessera Therapeutics, and Third Rock Ventures. M.A.H. consults for Akouos. J.M.R. consults for Maze Therapeutics.

The need for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) next-generation vaccines has been highlighted by the rise of variants of concern (VoCs) and the long-term threat of emerging coronaviruses. Here, we design and characterize four categories of engineered nanoparticle immunogens that recapitulate the structural and antigenic properties of the prefusion SARS-CoV-2 spike (S), S1, and receptor-binding domain (RBD). These immunogens induce robust S binding, ACE2 inhibition, and authentic and pseudovirus neutralizing antibodies against SARS-CoV-2. A spike-ferritin nanoparticle (SpFN) vaccine elicits neutralizing titers (ID 50 > 10,000) following a single immunization, whereas RBD-ferritin nanoparticle (RFN) immunogens elicit similar responses after two immunizations and also show durable and potent neutralization against circulating VoCs. Passive transfer of immunoglobulin G (IgG) purified from SpFN- or RFN-immunized mice protects K18-hACE2 transgenic mice from a lethal SARS-CoV-2 challenge. Furthermore, S-domain nanoparticle immunization elicits ACE2-blocking activity and ID 50 neutralizing antibody titers >2,000 against SARS-CoV-1, highlighting the broad response elicited by these immunogens., Competing Interests: Declaration of interests M.G.J. and K.M. are named as inventors on international patent application WO/2021/178971 A1 entitled “Vaccines against SARS-CoV-2 and other coronaviruses.” M.G.J. is named as an inventor on international patent application WO/2018/081318 and U.S. patent 10,960,070 entitled “Prefusion coronavirus spike proteins and their use.” Z.B. is named as an inventor on U.S. patent 10,434,167 entitled “Non-toxic adjuvant formulation comprising a monophosphoryl lipid A (MPLA)-containing liposome composition and a saponin.” Z.B. and G.R.M. are named as inventors on U.S. patent application 16/607,917 entitled “Compositions and methods for vaccine delivery.” M.S.D. is a consultant for Inbios, Vir Biotechnology, Fortress Biotech, and Carnival Corporation and on the scientific advisory boards of Moderna and Immunome. The Diamond laboratory has received funding support from sponsored research agreements from Moderna, Vir Biotechnology, Kaleido, and Emergent BioSolutions. S.R., P.M.M., and M.T.E. are employees of AstraZeneca and currently hold AstraZeneca stock or stock options. Z.B. is currently employed at Pfizer., (Copyright © 2021. Published by Elsevier Inc.)