Multivariate curve resolution-based data fusion approaches applied in 1H NMR metabolomic analysis of healthy cohorts.

BACKGROUND: Metabolomics plays a critical role in deciphering metabolic alterations within individuals, demanding the use of sophisticated analytical methodologies to navigate its intricate complexity. While many studies focus on single biofluid types, simultaneous analysis of multiple matrices enhances understanding of complex biological mechanisms. Consequently, the development of data fusion methods enabling multiblock analysis becomes essential for comprehensive insights into metabolic dynamics. RESULTS: This study introduces a novel guideline for jointly analyzing diverse metabolomic datasets (serum, urine, metadata) with a focus on metabolic differences between groups within a healthy cohort. The guideline presents two fusion strategies, 'Low-Level data fusion' (LLDF) and 'Mid-Level data fusion' (MLDF), employing a sequential application of Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS), linking the outcomes of successive analyses. MCR-ALS is a versatile method for analyzing mixed data, adaptable at various stages of data processing-encompassing resonance integration, data compression, and exploratory analysis. The LLDF and MLDF strategies were applied to 1H NMR spectral data extracted from urine and serum samples, coupled with biochemical metadata sourced from 145 healthy volunteers. SIGNIFICANCE: Both methodologies effectively integrated and analysed multiblock datasets, unveiling the inherent data structure and variables associated with discernible factors among healthy cohorts. While both approaches successfully detected sex-related differences, the MLDF strategy uniquely revealed components linked to age. By applying this analysis, we aim to enhance the interpretation of intricate biological mechanisms and uncover variations that may not be easily discernible through individual data analysis.

Microplastics in seafood: Consumer preferences and valuation for mitigation technologies.

Microplastics, an emerging pollutant, have garnered widespread attention due to potential repercussions on human health and the environment. Given the critical role of seafood in food security, growing concerns about microplastics might be detrimental to meeting future global food demand. This study employed a discrete choice experiment to investigate Chilean consumers' preferences for technology aimed at mitigating microplastic levels in mussels. Using a between-subjects design with information treatments, we examined the impact of informing consumers about potential human health and environmental effects linked to microplastics pollution on their valuation for the technology. We found that the information treatments increased consumers' willingness to pay for mussels. Specifically, consumers were willing to pay a premium of around US$ 4 for 250 g of mussel meat with a 90 % depuration efficiency certification. The provision of health impact information increased the price premium by 56 %, while the provision of environmental information increased it by 21 %. Furthermore, combined health and environmental information significantly increased the probability of non-purchasing behavior by 22.8 % and the risk perception of microplastics for human health by 5.8 %. These results emphasized the critical role of information in shaping consumer preferences and provided evidence for validating investment in research and development related to microplastic pollution mitigation measures.

The mental burden of stay-at-home order extensions during COVID-19.

This study evaluates the psychological impact of stay-at-home extension orders during COVID-19 and its relationship with individuals' expectations on the duration of the extensions. An online survey was administered to 1259 US adult residents to measure symptoms of post-traumatic stress disorder (PTSD), anxiety and stress induced by different stay-at-home order extensions using hypothetical length scenarios. We find that individuals exposed to two 2-week order extensions exhibit higher levels of stress and anxiety compared to those exposed to a single 4-week extension. We also find that subjects with longer expected extensions exhibit more signs of psychological damage than those with shorter expected extensions. Furthermore, we find that the negative psychological consequences of providing two shorter extensions is observed only among subjects with extension expectations of four weeks or less. Our results demonstrate that people's expectations affect the level of psychological damage caused by lockdown mandates. Our findings suggest that whenever lockdown extensions are necessary, reduced psychological distress may be possible by implementing a one-time restriction, rather than extending multiple smaller extensions perhaps due to manipulation of personal expectations.

Thermal Stabilization of a Bacterial Zn(II)-Dependent Phospholipase C through Consensus Sequence Design.

Proteins' extraordinary performance in recognition and catalysis has led to their use in a range of applications. However, proteins obtained from natural sources are oftentimes not suitable for direct use in industrial or diagnostic setups. Natural proteins, evolved to optimally perform a task in physiological conditions, usually lack the stability required to be used in harsher conditions. Therefore, the alteration of the stability of proteins is commonly pursued in protein engineering studies. Here, we achieved a substantial thermal stabilization of a bacterial Zn(II)-dependent phospholipase C by consensus sequence design. We retrieved and analyzed sequenced homologues from different sources, selecting a subset of examples for expression and characterization. A non-natural consensus sequence showed the highest stability and activity among those tested. Comparison of the stability parameters of this stabilized mutant and other natural variants bearing similar mutations allows us to pinpoint the sites most likely to be responsible for the enhancement. Point mutations in these sites alter the unfolding process of the consensus sequence. We show that the stabilized version of the protein retains full activity even in harsh oil degumming conditions, making it suitable for industrial applications.

Safety and Immunogenicity of an Andes Virus DNA Vaccine by Needle-Free Injection: A Randomized, Controlled Phase 1 Study.

BACKGROUND: Andes virus (ANDV), a rodent-borne hantavirus, causes hantavirus pulmonary syndrome (HPS). The safety and immunogenicity of a novel ANDV DNA vaccine was evaluated. METHODS: Phase 1, double-blind, dose-escalation trial randomly assigned 48 healthy adults to placebo or ANDV DNA vaccine delivered via needle-free jet injection. Cohorts 1 and 2 received 2 mg of DNA or placebo in a 3-dose (days 1, 29, 169) or 4-dose (days 1, 29, 57, 169) schedule, respectively. Cohorts 3 and 4 received 4 mg of DNA or placebo in the 3-dose and 4-dose schedule, respectively. Subjects were monitored for safety and neutralizing antibodies by pseudovirion neutralization assay (PsVNA50) and plaque reduction neutralization test (PRNT50). RESULTS: While 98% and 65% of subjects had at least 1 local or systemic solicited adverse event (AE), respectively, most AEs were mild or moderate; no related serious AEs were detected. Cohorts 2, 3, and 4 had higher seroconversion rates than cohort 1 and seropositivity of at least 80% by day 197, sustained through day 337. PsVNA50 geometric mean titers were highest for cohort 4 on and after day 197. CONCLUSIONS: This first-in-human candidate HPS vaccine trial demonstrated that an ANDV DNA vaccine was safe and induced a robust, durable immune response. Clinical Trials Registration. NCT03682107.

Report of the Assay Guidance Workshop on 3-Dimensional Tissue Models for Antiviral Drug Development.

The National Center for Advancing Translational Sciences (NCATS) Assay Guidance Manual (AGM) Workshop on 3D Tissue Models for Antiviral Drug Development, held virtually on 7-8 June 2022, provided comprehensive coverage of critical concepts intended to help scientists establish robust, reproducible, and scalable 3D tissue models to study viruses with pandemic potential. This workshop was organized by NCATS, the National Institute of Allergy and Infectious Diseases, and the Bill and Melinda Gates Foundation. During the workshop, scientific experts from academia, industry, and government provided an overview of 3D tissue models' utility and limitations, use of existing 3D tissue models for antiviral drug development, practical advice, best practices, and case studies about the application of available 3D tissue models to infectious disease modeling. This report includes a summary of each workshop session as well as a discussion of perspectives and challenges related to the use of 3D tissues in antiviral drug discovery.

Effect of saline intrusion on rice production in the Mekong River Delta.

Saline intrusion is increasingly threatening the rice farming system in The Mekong River Delta (MRD). Identifying the impact of this disaster on rice farming and providing promptly adaptable solutions is an urgent issue. This study evaluates the influence of saline intrusion on rice productivity of households in the MRD. We utilize the quasi-experimental method, the difference-in-difference (DID) method, with farm level panel data on rice farming extracted from the Vietnam Household Living Standards Survey (VHLSS) in 2014 and 2016. The empirical results indicate that households affected by saline intrusion have lower total production and lower productivity compared to the unaffected group. The affected group has lower total revenue of approximately 4969.8 thousand VND per ha and lower net revenue, on an average of 4679.3 thousand VND per ha, compared to the comparable unaffected group. Among the subsamples of different regions, the damage magnitude of households in severely affected regions by salinity intrusion is higher than in less severely affected areas. Some policy implications that can be proposed from this research are that the management agency should continue to update information on weather and climate change scenarios for forecasting and timely information for rice farmers. The agency comprehensively evaluates the effectiveness of transformed models of sustainable agricultural production adapted to salinity intrusion and replicates high-effective production models; restructure rice cultivation following the salinity intrusion level of each locality.

Sustainable Refining of Vegetable Oil Made Easy with a Designer Phospholipase C Enzyme.

The increasing demand pressures the vegetable oil industry to develop novel refining methods. Degumming with type C phospholipases (PLCs) is a green technology and provides extra oil. However, natural PLCs are not active under the harsh conditions used in oil refining plants, requiring additional unit operations. These upfront capital expenditures and the associated operational costs hinder the adoption of this method. Here, we present a process based on ChPLC, a synthetic PLC obtained by consensus sequence design, possessing superior thermal stability and catalytic properties. Using ChPLC, crude soybean oil degumming was completed at 80 °C in 30 min, the temperature and residence time imposed by the design of existing oil refining plants. Remarkably, an extra yield of oil of 2% was obtained using 60% of the dose recommended for PLCs marketed today, saving upfront investments and reducing the operational cost of degumming. A techno-economic analysis indicates that, for medium size plants, ChPLC reduces the overall cost of soybean oil enzymatic degumming by 58%. The process presented here facilitates the implementation of enzymatic technologies to oil producers, regardless of their processing capacity, bringing potential annual benefits in the billion-dollar range for the global economy.

Did mental and emotional health of SNAP families' children improve during the COVID-19 pandemic?

The onset of the COVID-19 pandemic disproportionately affected low-income households in the United States. As part of the government's response to the pandemic, households with children participating in the Supplemental Nutrition Assistance Program (SNAP) benefited from several temporary provisions. This study examines whether the mental/emotional well-being of children in SNAP families was influenced by the SNAP temporary provisions, overall and across subpopulations by race/ethnicity and school meal programs (SMP) participation status of children. Cross-sectional data from the 2016-2020 National Survey of Children Health (NSCH) were used to study the occurrence of mental, emotional, developmental, or behavioral (MEDB) health of children (6-17 years) in SNAP families. Difference-in-Differences (DID) analyses were conducted to assess the association between MEDB health of children in SNAP families and the implementation of the SNAP provisions. Overall, finding show that between 2016 and 2020, children in SNAP families were more likely to face adverse MEDB conditions than children in non-SNAP families (p < 0.05). DID analyses indicate that children's MEDB health was not statistically affected following the first year of SNAP temporary provisions (p > 0.1). Additionally, no differential results were found by race/ethnicity of children or SMP participation (p > 0.1). Results are robust to the use of different well-being measures. These results suggest that SNAP provisions may have been associated with the reduction of the adverse effects of the pandemic on children's well-being.

Relationship between different sources of non-protein nitrogen and supplementation times on performance and metabolism of grazing Nellore cattle during the dry season.

Two experiments were conducted to evaluate the effect of supplementation with two sources of non-protein nitrogen at different feeding times on the performance, ingestive behavior, and rumen metabolism of growing Nellore bulls during the dry season. Exp. 1: One hundred and twenty Nellore bulls, weighing 206 ± 39 kg of initial body weight (BW) and 12 months of age, were divided into 20 paddocks, and they were used in randomized block design in a 2 × 2 factorial arrangement to evaluate performance and ingestive behavior. Exp. 2: 12 rumen cannulated animals with 509 ± 59 BW, divided into 4 paddocks, were used in a triple Latin square 4 × 4 in a 2 × 2 factorial arrangement to evaluate metabolism. The factors were 2 non-protein nitrogen sources (urea or slow-release urea) and 2 feeding times (07:00 or 13:00 at 4 g/kg BW of supplement). There was no influence of non-protein sources, supplementation time, or their interaction on the grazing time or the trough time during daytime, nighttime, or total (P ≥ 0.16). There were no interactions or factor effects on ADG (P ≥ 0.45) or final body weight (P ≥ 0.39). There was an interaction between supplementation time and collection time (P < 0.01) on ruminal pH. Animals supplemented in the morning had greater total SCFA at 18 h after supplementation (P = 0.03). The supplementation time and the non-protein nitrogen sources did not alter the ingestive behavior or animal performance of young Nellore cattle.

COVID-19 Detection on Chest X-ray and CT Scan: A Review of the Top-100 Most Cited Papers.

Since the beginning of the COVID-19 pandemic, many works have been published proposing solutions to the problems that arose in this scenario. In this vein, one of the topics that attracted the most attention is the development of computer-based strategies to detect COVID-19 from thoracic medical imaging, such as chest X-ray (CXR) and computerized tomography scan (CT scan). By searching for works already published on this theme, we can easily find thousands of them. This is partly explained by the fact that the most severe worldwide pandemic emerged amid the technological advances recently achieved, and also considering the technical facilities to deal with the large amount of data produced in this context. Even though several of these works describe important advances, we cannot overlook the fact that others only use well-known methods and techniques without a more relevant and critical contribution. Hence, differentiating the works with the most relevant contributions is not a trivial task. The number of citations obtained by a paper is probably the most straightforward and intuitive way to verify its impact on the research community. Aiming to help researchers in this scenario, we present a review of the top-100 most cited papers in this field of investigation according to the Google Scholar search engine. We evaluate the distribution of the top-100 papers taking into account some important aspects, such as the type of medical imaging explored, learning settings, segmentation strategy, explainable artificial intelligence (XAI), and finally, the dataset and code availability.

A disordered region retains the full protease inhibitor activity and the capacity to induce CD8+ T cells in vivo of the oral vaccine adjuvant U-Omp19.

U-Omp19 is a bacterial protease inhibitor from Brucella abortus that inhibits gastrointestinal and lysosomal proteases, enhancing the half-life and immunogenicity of co-delivered antigens. U-Omp19 is a novel adjuvant that is in preclinical development with various vaccine candidates. However, the molecular mechanisms by which it exerts these functions and the structural elements responsible for these activities remain unknown. In this work, a structural, biochemical, and functional characterization of U-Omp19 is presented. Dynamic features of U-Omp19 in solution by NMR and the crystal structure of its C-terminal domain are described. The protein consists of a compact C-terminal beta-barrel domain and a flexible N-terminal domain. The latter domain behaves as an intrinsically disordered protein and retains the full protease inhibitor activity against pancreatic elastase, papain and pepsin. This domain also retains the capacity to induce CD8+ T cells in vivo of U-Omp19. This information may lead to future rationale vaccine designs using U-Omp19 as an adjuvant to deliver other proteins or peptides in oral formulations against infectious diseases, as well as to design strategies to incorporate modifications in its structure that may improve its adjuvanticity.

Characterization of Otolaryngology Referrals among Pacific Islanders in the Commonwealth of the Northern Mariana Islands.

The burden of otolaryngology disease in Pacific Islander populations is relatively uncharacterized. A single-institution retrospective review was undertaken at the Commonwealth Healthcare Corporation in Saipan, the only hospital in the Commonwealth of the Northern Mariana Islands. Demographic, diagnostic, and treatment data were compiled from the clinical charts of all patients seen by an otolaryngologist between January 2015 and April 2020. For all Pacific Islanders in the sample (N=674), the average age was 40.2 (SD 22.4) years and ages ranged from 10 months to 89 years. Patients were 50.7% male and 49.3% female. The most common diagnoses affected the ear (40.8%), followed by the oral cavity/pharynx (23.2%), and nose (14.0%). Middle ear disease comprised 41.7% of reported ear disorders; the most common problem was otitis media (19.4%, n=68) followed by tympanic membrane perforation (14.0%, n=49). Head and neck cancers comprised 8.6% of all diagnoses. Most (77.8%) malignant neoplasms were oral cavity carcinomas. The average age at diagnosis for oral cancer was 46.6 years with a 1.8:1 male-to-female predominance. Patients with cancer of the oral cavity (n=56) chewed betel nut at higher rates (94.6%) compared with other adults in the sample (P<.001). Adult patients reported alcohol use, smoking, and chewing betel quid at rates of 26.5%, 39.9%, and 52.2% respectively. Otolaryngology referrals among Pacific Islanders in this sample were dominated by ear disease and included betel nut-related oral cavity disease.

US workers' willingness to accept meatpacking jobs amid the COVID-19 pandemic.

We implement a discrete choice experiment to examine the impact of COVID-19 exposure risk, unemployment risk, enhanced and extended unemployment benefits, and job attributes on low-skilled workers' willingness to accept (WTA) meatpacking jobs. With a sample average WTA wage of $22.77/h, the current national average meatpacking wage of approximately $15/h is too low for these workers to consider this employment opportunity. Enhanced layoff risk and exposure to COVID-19 further deterred respondents, while health insurance, retirement benefits, and a signing bonus enhanced respondents' WTA. The additional unemployment benefits of the CARES Act neither deterred nor encouraged respondents WTA.

Ezrin and CD44 participate in the internalization process of Coxiella burnetii into non-phagocytic cells.

Ezrin protein is involved in the interaction of actin cytoskeleton with membrane receptors such as CD44. It regulates plasma membrane dynamics and intracellular signaling. Coxiella burnetii, the etiologic agent of Q fever, is internalized into host cell through a poorly characterized molecular mechanism. Here we analyzed the role of ezrin and CD44 in the C. burnetii internalization by HeLa cells. The knockdown of ezrin and CD44 inhibited the bacterial uptake. Interestingly, at early stages of C. burnetii internalization, ezrin was recruited to the cell membrane fraction and phosphorylated. Moreover, the overexpression of non-phosphorylatable and phosphomimetic ezrin mutants decreased and increased the bacterial entry, respectively. A decrease in the internalization of C. burnetii was observed by the overexpression of CD44 truncated forms containing the intracellular or the extracellular domains. Interestingly, the CD44 mutant was unable to interact with ERM proteins decreased the bacterial internalization. These findings demonstrate the participation of ezrin in the internalization process of C. burnetii in non-phagocytic cells. Additionally, we present evidence that CD44 receptor would be involved in that process.

Characterisation of kinetics, substrate inhibition and product activation by AMP of bifunctional ADP-dependent glucokinase/phosphofructokinase from Methanococcus maripaludis.

Methanogenic archaea have received attention due to their potential use in biotechnological applications such as methane production, so their metabolism and regulation are topics of special interest. When growing in a nutrient-rich medium, these organisms exhibit gluconeogenic metabolism; however, under starvation conditions, they turn to glycolytic metabolism. To date, no regulatory mechanism has been described for this gluconeogenic/glycolytic metabolic switch. Here, we report that adenosine monophosphate (AMP) activates both enzymatic activities of the bifunctional adenosine diphosphate (ADP)-dependent phosphofructokinase/glucokinase from Methanococcus maripaludis (MmPFK/GK). To understand this phenomenon, we performed a comprehensive kinetic characterisation, including determination of the kinetics, substrate inhibition and AMP activation mechanism of this enzyme. We determined that MmPFK/GK has an ordered-sequential mechanism, in which MgADP is the first substrate to bind and AMP is the last product released. The enzyme also displays substrate inhibition by both sugar substrates; we determined that this inhibition occurs through the formation of catalytically nonproductive enzyme complexes caused by sugar binding. For both activities, the AMP activation mechanism occurs primarily through incremental changes in the affinity for the sugar substrate, with this effect being higher in the GK than in the PFK activity. Interestingly, due to the increase in the sugar substrate affinity caused by AMP, an enhancement in the sugar substrate inhibition effect was also observed for both activities, which can be explained by an increase in sugar binding leading to the formation of dead-end complexes. These results shed light on the regulatory mechanisms of methanogenic archaeal sugar metabolism, a phenomenon that has been largely unexplored.

Health versus income amid COVID-19: What do people value more?

Public efforts to battle COVID-19 have been portrayed as a trade-off between health and the economy in the U.S. public discourse. We investigate how the U.S. general public prioritizes the health and the income dimensions amid COVID-19 using an incentivized instrument with real monetary consequences. We also employ between-subject information treatments highlighting negative health and income consequences of the pandemic. Specifically, participants have to divide monetary contributions between two charitable organizations representing either the health or the income dimension. An overwhelming majority of participants supports both dimensions, with higher monetary contributions to the health dimension (56%) compared to income (44%), but the difference is not large. Only a small fraction of respondents contributes exclusively to the health (10%) or income (5%) dimensions. Increasing the salience of negative health outcomes of COVID-19 raises differential token allocations in favor of the health-oriented charity. This finding is important since the course of COVID-19 will be shaped by the policies governments implement and how the general public reacts to these policies.

Evaluation of Delivering Breakfast After the Bell and Academic Performance Among Third-Grade Children: An Application of the Synthetic Control Method.

BACKGROUND: Breakfast after the bell (BAB), an alternative way to deliver breakfast after the school day begins, has been shown to increase participation in the School Breakfast Program. However, BAB occupies time that could otherwise be used for instruction and may affect academic performance. This study examined whether BAB affects math and literacy scores in third grade, an age not adequately studied in earlier literature. METHODS: Using data on Arkansas students, we compared schools adopting BAB to corresponding "synthetic" control schools by minimizing preadoption differences in observables that can influence test scores. RESULTS: We found little evidence of positive or negative effects on test scores. We also found little evidence of meaningful program effects over subsamples of schools by district enrollment size and BAB delivery method. Schools that adopted grab-and-go delivery and schools in districts with small enrollments, less than 600 students, showed statistically negative effects on math after BAB adoption. However, these effects were transitory and no longer present by the time the first fully exposed cohort reached third grade. CONCLUSIONS: BAB can be incorporated into the school day without adversely impacting academic achievement, especially if adopting schools seek input from educators and nutrition personnel already experienced with BAB.

Impact of Weight Status Reporting on Childhood Body Mass Index.

Background: Many states have adopted school-based BMI screening or surveillance programs in an effort to address high rates of childhood obesity, some of which involve provision of confidential BMI reports to parents. While there is evidence that parents are attuned to information in the reports, there is less evidence showing that the reports are effective in preventing excess childhood weight gain. Methods: Data from Arkansas, the state with the nation's first and longest running and BMI screening program, were used to measure the impact of BMI reports. This was done through a regression discontinuity design that compared future BMI z-scores among children falling within a narrow band around the obese and overweight thresholds. We derived the effects of BMI reports by comparing students who received different types of reports around the relevant threshold. Results: While we are unable to detect any differences in BMI z-scores between the children who received the overweight report and the children who received the healthy weight report, we detected some differences between children who received the obese report and children who received the overweight report. These findings hold across subsamples by age, minority status, and school meal status. Conclusions: Based on these data, overweight or obese reports to do not meaningfully impact future BMI z-scores. This may be due, in part, to the format of parental reports, which may dampen the surprise element of an overweight or obese report.

Melanoma in children and adolescents: analysis of susceptibility genes in 123 Italian patients.

BACKGROUND: A polygenic inheritance involving high, medium and low penetrance genes has been suggested for melanoma susceptibility in adults, but genetic information is scarce for paediatric patients. OBJECTIVE: We aim to analyse the major high and intermediate melanoma risk genes, CDKN2A, CDK4, POT1, MITF and MC1R, in a large multicentre cohort of Italian children and adolescents in order to explore the genetic context of paediatric melanoma and to reveal potential differences in heritability between children and adolescents. METHODS: One-hundred-twenty-three patients (<21 years) from nine Italian centres were analysed for the CDKN2A, CDK4, POT1, MITF, and MC1R melanoma predisposing genes. The rate of gene variants was compared between sporadic, familial and multiple melanoma patients and between children and adolescents, and their association with clinico-pathological characteristics was evaluated. RESULTS: Most patients carried MC1R variants (67%), while CDKN2A pathogenic variants were found in 9% of the cases, the MITF E318K in 2% of patients and none carried CDK4 or the POT1 S270N pathogenic variant. Sporadic melanoma patients significantly differed from familial and multiple cases for the young age at diagnosis, infrequent red hair colour, low number of nevi, low frequency of CDKN2A pathogenic variants and of the MC1R R160W variant. Melanoma in children (≤12 years) had more frequently spitzoid histotype, were located on the head/neck and upper limbs and had higher Breslow thickness. The MC1R V92M variant was more common in children than in adolescents. CDKN2A common polymorphisms and MC1R variants were associated with a high number of nevi. CONCLUSION: Our results confirm the scarce involvement of the major high-risk susceptibility genes in paediatric melanoma and suggest the implication of MC1R gene variants especially in the children population.