Confinement plus myosin-II suppression maximizes heritable loss of chromosomes, as revealed by live-cell ChReporters.

The mechanical environment of a cell can have many effects, but whether it impacts the DNA sequence of a cell has remained unexamined. To investigate this, we developed a live-cell method to measure changes in chromosome numbers. We edited constitutive genes with GFP or RFP tags on single alleles and discovered that cells that lose Chromosome reporters (ChReporters) become non-fluorescent. We applied our new tools to confined mitosis and to inhibition of the putative tumor suppressor myosin-II. We quantified compression of mitotic chromatin in vivo and demonstrated that similar compression in vitro resulted in cell death, but also rare and heritable ChReptorter loss. Myosin-II suppression rescued lethal multipolar divisions and maximized ChReporter loss during three-dimensional (3D) compression and two-dimensional (2D) lateral confinement, but not in standard 2D culture. ChReporter loss was associated with chromosome mis-segregation, rather than just the number of divisions, and loss in vitro and in mice was selected against in subsequent 2D cultures. Inhibition of the spindle assembly checkpoint (SAC) caused ChReporter loss in 2D culture, as expected, but not during 3D compression, suggesting a SAC perturbation. Thus, ChReporters enable diverse studies of viable genetic changes, and show that confinement and myosin-II affect DNA sequence and mechano-evolution.

Where is the baby in core knowledge?

What we know about what babies know - as represented by the core knowledge proposal - is perhaps missing a place for the baby itself. By studying the baby as an actor rather than an observer, we can better understand the origins of human intelligence as an interface between perception and action, and how humans think and learn about themselves in a complex world.

Adaptation of intestinal fermentation over time in the growing pig is influenced by the amount of kiwi fruit consumed.

The effect of kiwi fruit at two dietary levels on the adaptation of intestinal fermentation over time in the growing pig was studied. A semi-synthetic fibre-free diet and two semi-synthetic diets containing kiwi fruit as a model fibre source (133 or 266 g/kg (DM basis); 28 or 48 g fibre/kg) were formulated and the diets contained titanium dioxide as an indigestible marker. A total of fourteen ileal cannulated pigs (41 kg body weight) were fed the fibre-free diet for 7 d followed by either the low or high kiwi fruit-containing diets (n 7/diet) for a further 44 d. Ileal digesta and faeces were collected at five times throughout the study. Ileal digesta were fermented (in vitro) with a standard pooled human faecal inoculum, while fresh pig faeces were used as inocula to ferment in vitro a standard purified fibre. Observations were normalised for diet DM intake using the marker. The 16S ribosomal RNA gene copy number of ileal and total faecal bacteria were high for the high-kiwi fruit level diet (P<0·05). The ileal bacteria tended to decrease over time (P<0·1), while the faecal bacteria increased (P<0·05), at the same rate for both diets. The amounts of crude protein and insoluble dietary fibre entering the hindgut changed over time similarly for both diets, whereas for starch it changed only for the low kiwi fruit-containing diet (P<0·05). Changes over time were also observed for the predicted hindgut valeric acid production and butyric acid absorption (P<0·05). In conclusion, adaptational changes over time of some characteristics of intestinal fermentation depended on the dietary level of kiwi fruit.

Differences in cell shape, motility, and growth reflect chromosomal number variations that can be visualized with live-cell ChReporters.

Chromosome numbers often change dynamically in tumors and cultured cells, which complicates therapy as well as understanding genotype-mechanotype relationships. Here we use a live-cell "ChReporter" method to identify cells with a single chromosomal loss in efforts to better understand differences in cell shape, motility, and growth. We focus on a standard cancer line and first show clonal populations that retain the ChReporter exhibit large differences in cell and nuclear morphology as well as motility. Phenotype metrics follow simple rules, including migratory persistence scaling with speed, and cytoskeletal differences are evident from drug responses, imaging, and single-cell RNA sequencing. However, mechanotype-genotype relationships between fluorescent ChReporter-positive clones proved complex and motivated comparisons of clones that differ only in loss or retention of a Chromosome-5 ChReporter. When lost, fluorescence-null cells show low expression of Chromosome-5 genes, including a key tumor suppressor APC that regulates microtubules and proliferation. Colonies are compact, nuclei are rounded, and cells proliferate more, with drug results implicating APC, and patient survival data indicating an association in multiple tumor-types. Visual identification of genotype with ChReporters can thus help clarify mechanotype and mechano-evolution.

Gastric digestion of cow milk, almond milk and oat milk in rats.

In this study, gastric digestion of isocaloric and iso-macronutrient cow milk, almond milk and oat milk were compared in rats euthanized at different post-feeding times. The cow milk separated into a curd phase and a liquid phase in the rat stomach. This coagulation of the cow milk led to higher (P < 0.05) protein and lipid retention in the stomach compared with almond milk and oat milk. Almond milk oil bodies aggregated, creamed and rapidly layered in the stomach. This induced a faster (P < 0.05) gastric emptying of proteins (T1/2 = 36 min) compared with cow milk (T1/2 = 89 min) and oat milk (T1/2 = 55 min), and a slower gastric emptying of almond lipids than of almond proteins. In contrast, no significant physical change during the digestion of oat milk was found, with both the proteins and the lipids being steadily emptied from the stomach. This in vivo study provides information on the gastric digestion and emptying (and thereby nutritional characteristics) of plant-based milks compared with animal-based milks, that will be useful for the design of novel plant-based drinks.

A digital approach to asthma self-management in adults: Protocol for a pragmatic randomized controlled trial.

Asthma self-management can improve symptom control, but adherence to established self-management behaviors is often poor. With adult asthma uncontrolled in over 60% of U.S. cases, there is a need for scalable, cost-effective tools to improve asthma outcomes. Here we describe a protocol for the Asthma Digital Study, a 24-month, decentralized, pragmatic, open-label, randomized controlled trial investigating the impact of a digital asthma self-management (DASM) program on asthma outcomes in adults. The program leverages consumer-grade devices with a smartphone app to provide "smart nudges," symptom logging, trigger tracking, and other features. Participants are recruited (target N = 900) from throughout the U.S., and randomized to a DASM or control arm (1:1). Co-primary outcomes at one year are a) asthma-associated costs for acute care and b) change from baseline in Asthma Control Test™ scores. Findings may inform decisions around adoption of digital tools for asthma self-management. Trial registration:clinicaltrials.gov identifier: NCT04609644. Registered: Oct 30, 2020.

Salmon food matrix influences digestion and bioavailability of long-chain omega-3 polyunsaturated fatty acids.

The natural structure of whole food plays an important role in the physiological impact of bioactive compounds present within the food, also known as the "matrix effect". Long-chain omega-3 polyunsaturated fatty acids (LCn-3PUFAs) are one example of a food-derived nutrient, mostly found in fish, that is believed to be influenced by the food matrix. However, most previous studies have compared only the long-term bioavailability of fish versus fish oil and have used commercial sources of fish oil. The present study aimed to investigate whether fish (salmon) matrix influences the transit of LCn-3PUFAs during in vitro digestion and affects bioavailability in healthy females. Meals containing intact salmon (intact structure), minced salmon (some structure) and defatted salmon + oil (no structure) with identical macronutrient compositions were developed. Healthy female participants (n = 13) consumed the meals in a postprandial crossover study and blood was collected at regular time points for 6 h post meal consumption. In parallel, in vitro digestion of the meals was performed using a human gastric simulator (HGS) and digesta samples were collected at regular time points for 6 h. Results: showed that plasma concentration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were significantly higher after participants consumed intact salmon compared to the other meals (covariate analysis p < 0.001). The in vitro digestion results showed defatted salmon + oil meal had a faster decrease in pH and faster fat emptying from the HGS than the other two meals. The defatted salmon + oil meal more closely followed fat emptying of a homogeneous unstructured meal, whereas the other meals exhibited phase separation with a delay in fat emptying. Conclusion: The fish matrix (salmon) plays an important role in the bioaccessibility and bioavailability of EPA and DHA. The differences are partly explained by fat digestion and emptying from the stomach. This study suggests that the natural structure of fish has a functional effect on the absorption and bioavailability of fish oil.

Latent Lyme Disease Resulting in Chronic Arthritis and Early Career Termination in a United States Army Officer.

Lyme disease is a continuing threat to military personnel operating in arboriferous and mountainous environments. Here we present the case of a 24-year-old Second Lieutenant, a recent graduate from the United States Military Academy, with a history of Lyme disease who developed recurrent knee effusions following surgery to correct a hip impingement. Although gonococcal arthritis was initially suspected from preliminary laboratory results, a comprehensive evaluation contradicted this diagnosis. Despite antibiotic therapy, aspiration of the effusions, and steroid treatment to control inflammation, the condition of the patient deteriorated to the point where he was found to be unfit for duty and subsequently discharged from active military service. This case illustrates the profound effect that latent Lyme disease can have on the quality of life and the career of an active duty military member. It highlights the need for increased surveillance for Borrelia burgdorferi (B. burgdorferi) in military training areas and for the early and aggressive diagnosis and treatment of military personnel who present with the symptoms of acute Lyme disease.