Prior Heterologous Flavivirus Exposure Results in Reduced Pathogenesis in a Mouse Model of Zika Virus Infection.

The 2015/2016 Zika virus epidemic in South and Central America left the scientific community urgently trying to understand the factors that contribute to Zika virus pathogenesis. Because multiple other flaviviruses are endemic in areas where Zika virus emerged, it is hypothesized that a key to understanding Zika virus disease severity is to study Zika virus infection in the context of prior flavivirus exposure. Human and animal studies have highlighted the idea that having been previously exposed to a different flavivirus may modulate the immune response to Zika virus. However, it is still unclear how prior flavivirus exposure impacts Zika viral burden and disease. In this murine study, we longitudinally examine multiple factors involved in Zika disease, linking viral burden with increased neurological disease severity, weight loss, and inflammation. We show that prior heterologous flavivirus exposure with dengue virus type 2 or 3 or the vaccine strain of yellow fever provides protection from mortality in a lethal Zika virus challenge. However, reduction in viral burden and Zika disease varies depending on the infecting primary flavivirus; with primary Zika virus infection being most protective from Zika virus challenge, followed by dengue virus 2, with yellow fever and dengue virus 3 protecting against mortality but showing more severe disease. This study demonstrates the variation in protective effects of prior flavivirus exposure on Zika virus pathogenesis and identifies distinct relationships between primary flavivirus infection and the potential for Zika virus disease. IMPORTANCE The emergence and reemergence of various vector-borne diseases in recent years highlights the need to understand the mechanisms of protection for each pathogen. In this study, we investigated the impact of prior exposure to Zika virus, dengue virus serotypes 2 or 3, or the vaccine strain of yellow fever on pathogenesis and disease outcomes in a mouse model of Zika virus infection. We found that prior exposure to a heterologous flavivirus was protective from mortality, and to varying degrees, prior flavivirus exposure was protective against neurological disease, weight loss, and severe viral burden during a lethal Zika challenge. Using a longitudinal and cross-sectional study design, we were able to link multiple disease parameters, including viral burden, with neurological disease severity, weight loss, and inflammatory response in the context of flavivirus infection. This study demonstrates a measurable but varied impact of prior flavivirus exposure in modulating flavivirus pathophysiology. Given the cyclic nature of most flavivirus outbreaks, this work will contribute to the forecasting of disease severity for future outbreaks.

Root nodules of red alder (Alnus rubra) and sitka alder (Alnus viridis ssp. sinuata) are inhabited by taxonomically diverse cultivable microbial endophytes.

The root nodules of actinorhizal plants are home to nitrogen-fixing bacterial symbionts, known as Frankia, along with a small percentage of other microorganisms. These include fungal endophytes and non-Frankia bacteria. The taxonomic and functional diversity of the microbial consortia within these root nodules is not well understood. In this study, we surveyed and analyzed the cultivable, non-Frankia fungal and bacterial endophytes of root nodules from red and Sitka alder trees that grow together. We examined their taxonomic diversity, co-occurrence, differences between hosts, and potential functional roles. For the first time, we are reporting numerous fungal endophytes of alder root nodules. These include Sporothrix guttuliformis, Fontanospora sp., Cadophora melinii, an unclassified Cadophora, Ilyonectria destructans, an unclassified Gibberella, Nectria ramulariae, an unclassified Trichoderma, Mycosphaerella tassiana, an unclassified Talaromyces, Coniochaeta sp., and Sistotrema brinkmanii. We are also reporting several bacterial genera for the first time: Collimonas, Psychrobacillus, and Phyllobacterium. Additionally, we are reporting the genus Serratia for the second time, with the first report having been recently published in 2023. Pseudomonas was the most frequently isolated bacterial genus and was found to co-inhabit individual nodules with both fungi and bacteria. We found that the communities of fungal endophytes differed by host species, while the communities of bacterial endophytes did not.

Merging Metabolic Modeling and Imaging for Screening Therapeutic Targets in Colorectal Cancer.

Cancer-associated fibroblasts (CAFs) play a key role in metabolic reprogramming and are well-established contributors to drug resistance in colorectal cancer (CRC). To exploit this metabolic crosstalk, we integrated a systems biology approach that identified key metabolic targets in a data-driven method and validated them experimentally. This process involved high-throughput computational screening to investigate the effects of enzyme perturbations predicted by a computational model of CRC metabolism to understand system-wide effects efficiently. Our results highlighted hexokinase (HK) as one of the crucial targets, which subsequently became our focus for experimental validation using patient-derived tumor organoids (PDTOs). Through metabolic imaging and viability assays, we found that PDTOs cultured in CAF conditioned media exhibited increased sensitivity to HK inhibition. Our approach emphasizes the critical role of integrating computational and experimental techniques in exploring and exploiting CRC-CAF crosstalk.

Analysis of estimation methods for resting metabolic rate in critically ill adults.

BACKGROUND: Prediction of metabolic rate is an important part of the nutrition assessment of critically ill patients, yet there are limited data regarding the best equation to use to make this prediction. METHODS: Standardized indirect calorimetry measurements were made in 202 ventilated, adult critical care patients, and resting metabolic rate was calculated using the following equations: Penn State equation, Faisy, Brandi, Swinamer, Ireton-Jones, Mifflin, Mifflinx1.25, Harris Benedict, Harris Benedictx1.25, Harris Benedict using adjusted weight for obesity, and each of the adjusted weight versions of Harris Benedictx1.25. The subjects were subgrouped by age and obesity status (young nonobese, young obese, elderly nonobese, elderly obese). Performance of each equation was assessed using bias, precision, and accuracy rate statistics. RESULTS: Accuracy rates in the study population ranged from 67% for the Penn State equation to 18% for the weight-adjusted Harris Benedict equation (without multiplication). Within subgroups, the highest accuracy rate was 77% in the elderly nonobese using the Penn State equation and the lowest was 0% for the weight-adjusted Harris Benedict equation. The Penn State equation was the only equation that was unbiased and precise across all subgroups. The obese elderly group was the most difficult to predict. Therefore, a separate regression was computed for this group: Mifflin(0.71)+Tmax(85)+Ve(64)-3085. CONCLUSIONS: The Penn State equation provides the most accurate assessment of metabolic rate in critically ill patients if indirect calorimetry is unavailable. An alternate form of this equation for elderly obese patients is presented, but has yet to be validated.

An evaluation of a handheld indirect calorimeter against a standard calorimeter in obese and nonobese adults.

BACKGROUND: Handheld indirect calorimetry has the potential to allow simple and inexpensive measurement of resting metabolic rate in spontaneously breathing people. However, validation work on these devices is contradictory. The purpose of the current study was to determine the bias and level of agreement of oxygen consumption and resting metabolic rate as measured by a handheld indirect calorimeter against a standard open-circuit indirect calorimetry cart. MATERIALS AND METHODS: One hundred community-living, spontaneously breathing, ambulatory nonobese and obese adults were studied in single sessions by a single investigator. Sequential measurements were undertaken using the handheld indirect calorimeter and the standard metabolic cart. Measurement sequence was varied randomly. RESULTS: The mean value for oxygen consumption and metabolic rate of the 2 devices was not significantly different. However, agreement between the 2 devices was only 43% in nonobese and obese participants, and there was proportional and fixed bias, with the handheld calorimeter tending to produce a higher value for oxygen consumption and resting metabolic rate. Limits of agreement for resting metabolic rate between the 2 calorimeters were -240 to +300 kcal/d. CONCLUSIONS: Measurements of resting metabolic rate by the handheld indirect calorimeter tested in this study are not equivalent to measurements by standard indirect calorimetry.

Recovery to resting metabolic state after walking.

Metabolic rate is usually measured in a resting state. To achieve this, a period of up to 30 minutes is given to recover from walking prior to the test. A work group from the American Dietetic Association recommends that 10 to 20 minutes is sufficient to achieve rest, but supporting data are limited. The purpose of this prospective observational study then was to determine how much time is needed for adults to recover to rest after walking 300 meters. Each participant's metabolic rate was measured with indirect calorimetry for 30 minutes after a 30-minute rest. The participant then walked 300 meters on a measured course, and metabolic rate was measured again for 30 minutes. Recovery to rest was considered to have occurred when the measured metabolic rate returned to a level of less than 6% above the resting measurement. Forty healthy ambulatory adults completed this study. Analysis of variance indicated that after a 300-meter walk, resting level of metabolic rate was achieved by the 10th minute of rest. However, it took 20 minutes for 95% of all participants to meet the 6% threshold (the remaining 5% who did not reach the threshold were observed to be moving during the measurement). The results of this study indicate that if a person lies still, recovery to rest after walking occurs by 20 minutes, validating the recommendation made by the expert panel of the American Dietetic Association's work group on indirect calorimetry. Rest periods of 30 minutes are not required, but the person should be observed for movement.