Dietary isoleucine restriction improves metabolic health and lifespan in animal models. It is hypothesized that shifts in branched-chain amino acid catabolism drive longevity. We investigated dietary isoleucine restriction on organismal branched-chain amino acid catabolism and its association with longevity and reproduction. In two main experiments, adult female Lubber grasshoppers were assigned to a high-quality diet (Bal ile) or diets with two levels of isoleucine content (moderate isoleucine restriction [Mod ile-R] or severe isoleucine restriction [Sev ile-R]). In Experiment 2, we used a low-energy (Low Lett) group as a positive control for catabolism. In Experiment 3, we used an ad libitum lettuce (Ad Lib Lett) control to show the reproduction-longevity trade-off. Nutrient-specific, organismal catabolism was measured by tracking [13]C in an ingested amino acid through catabolism to breath CO2. The Bal ile, Mod ile-R, and Sev ile-R diets extended lifespan and increased leucine catabolism relative to the Ad Lib Lett group. The Sev ile-R and Low Lett groups showed the highest leucine catabolism but greatly reduced reproduction. The Bal ile group showed intermediate leucine catabolism while sustaining reproduction. The Mod ile-R group had intermediate leucine catabolism but delayed reproduction. There was no compensatory feeding on low isoleucine diets. Valine catabolism did not show statistical significance but tended to show similar trends to leucine catabolism. Weak leucine catabolism is linked to a short lifespan, while excessive leucine catabolism is linked to impaired reproduction. Intermediate levels of leucine catabolism were associated with longevity and sustained reproduction.

We studied lifespan and lifetime reproductive success (LRS) in mice from a selection experiment that includes 4 replicate High Runner (HR) lines bred for voluntary wheel-running behavior and 4 non-selected Control (C) lines. HR mice typically run ~ threefold farther on a daily basis, leading to substantially increased energy expenditure and food consumption, and are also more active and eat more when housed without wheels. HR and C mice have been shown to differ for many other traits that could affect reproduction, including body mass and composition, aerobic capacity, hormonal profile (including corticosterone and leptin), and other behavioral traits (e.g., maternal care, thermoregulatory nesting, anxiety-like behavior in an elevated plus maze). At generation 97, male-female pairs were maintained until end of life in standard cages without wheels, with ad lib food and water. The number of pups weaned (LRS) ranged from 0-80, with a mean of 29.1 ± 8.1 (SE) for C lines and 25.8 ± 4.4 for HR (P = 0.73). Thus, artificial selection for high voluntary exercise did not significantly alter LRS. In addition, we found no statistical difference in average lifespan for dams or sires. However, replicate lines differed significantly for several traits, presumably due primarily to random genetic drift. At the level of individual variation, correlations among reproductive traits and lifespan provided little evidence for negative associations, and LRS was not negatively related to parental longevity. Overall, long-term selection on voluntary locomotor behavior did not produce detectable reductions in reproductive output or lifespan under standard laboratory conditions.

The oral delivery of biologics such as oligonucleotides may be enabled by the use of a chemical permeation enhancer such as sodium caprate (C10). However, C10 has low solubility under acidic gastric conditions which, often combined with poor drug stability, limits absorption when formulated in oral formulations. In this work, we investigated the use of alkalizers in immediate release C10-based formulations to enable the delivery of an oligonucleotide drug, RO7062931. Across the physiological pH range, C10 demonstrated low solubility and RO7062931 demonstrated poor chemical stability at pH values lower than 5, which together present significant obstacles to the oral delivery of the oligonucleotide. Screening alkalizers revealed that these pH-modifying components rapidly raised both the simulated gastric fluid bulk pH and microenvironmental pH when combined with C10, increasing the amount of C10 dissolving. Incorporating the most promising alkalizer, sodium carbonate, into C10-containing tablets of RO7062931 led to improved oligonucleotide chemical stability and C10 solubility under gastric conditions during in vitro testing, with the most effective formulation exhibiting complete RO7062931 stability and full C10 release. Alkalizer addition further contributed to oligonucleotide/C10 co-release, which is important for maximizing permeation enhancement activity at epithelial barriers. These findings offer insight into alkalizer incorporation as a promising strategy to improve the gastric delivery of oral formulations of oligonucleotide drugs and permeation enhancers.

Greenwashing tactics involve advertising products as ecofriendly and/or natural, which may reduce perceived health risk. We tested the effect of greenwashing advertising tactics on cigarette demand, puff topography, and perceived health risk. Young adults (N = 31) who smoke cigarettes completed a within-subject experiment involving two topography and seven operant demand sessions. Participants were exposed to advertising for novel cigarette "brands." One brand was advertised using greenwashing tactics, whereas the other brand included matched control tactics. Unknown to participants, both conditions used the same commercial cigarette brand. During topography sessions, participants smoked cigarettes ad lib and puff topography data were collected. During demand sessions, cigarettes were concurrently available at varying prices. No significant differences were observed for puff-topography measures (dz values < 0.28). Participants demonstrated a preference for the cigarettes advertised using greenwashing tactics when concurrently available at the same low price with more puffs earned (dz = 0.44) and a greater proportion of total puffs smoked (dz = 0.55). Both brands acted as substitutes for each other, with substitution not reliably differing by brand. These findings indicate that greenwashing advertising tactics may increase the relative reinforcing effects of cigarettes particularly when available at prices similar to those for standard cigarettes, findings that should be considered by regulatory authorities.

Bed- and risetimes of human ad lib sleep are determined by the endogenous mechanism of sleep-wake regulation. However, humans are capable to volitionally prolong (i.e. without any external influences) the wake phase of their sleep-wake cycle to early night hours. We hypothesized that the conscious behaviors mostly known under the terms "bedtime procrastination" and "mind-bedtime procrastination" can compete with the biological sleep-wake regulator for setting bedtimes on weekdays and/or weekends and thus this behavior can contribute to the delay of sleep relative to the bedtime signal from this regulator. University students (5375 in total) were grouped in accord with their earlier or later weekday and weekend bedtimes. To test the hypothesis, the group-averaged sleep times were calculated and simulated. Near 8% of students reported later weekday than weekend bedtimes. The simulations of their sleep times suggested that these times cannot be explained by a model of sleep-wake regulator without the additional assumption of bedtime delay caused by the volitional missing the bedtime signal from this regulator (i.e. this missing can be a result of such volitional behavior as mind-bedtime procrastination on weekdays rather than on weekends). We concluded that the concept of two conflicting - biological and "social" - "clocks" that was introduced in this journal 20 years ago can be extended to the concept of three conflicting "clocks." Bed- and risetimes on weekdays and weekends can reflect the competition between three - biological, "social," and, additionally, "mental" - "clocks." In accord with the concept of two "conflicting clocks," weekday risetimes can be mostly determined by "social clocks" competing with the biological clocks that can be solely responsible for weekend risetimes. However, weekday and/or weekend bedtimes cannot be always set by the biological clocks. Sometimes, they are determined by the additional conflict of "mental (mind) clocks" with the biological (body) clocks. This conflict can lead to weekday sleep insufficiency and can be, in particular, explained by such kind of conscious behavior as mind-bedtime procrastination.

Electroencephalography (EEG) signal is driven by oscillations arising from the synchronized activity of cortical neuronal populations. Phase-amplitude coupling (PAC) refers to the interaction whereby the amplitude of higher-frequency oscillations is systematically modulated by the phase of slower rhythms. It has been suggested that the excitation/inhibition (E/I) balance, which is often disrupted in neuropsychiatric and neurodegenerative disorders, plays a critical role in shaping PAC, although direct evidence in humans remains limited. In this study, we investigated the effects of pharmacologically modulated E/I balance on PAC using resting-state EEG (rs-EEG) data. Specifically, we analyzed the impact of anti-glutamatergic drugs, dextromethorphan (NMDA receptor antagonist) and perampanel (AMPA receptor antagonist), as well as nimodipine (L-type voltage-gated calcium channel blocker) on PAC in a double blind, placebo-controlled experiment. Analysis revealed a significant decrease in PAC following administration of dextromethorphan and perampanel, while nimodipine and placebo showed no effects. By showing that PAC is sensitive to pharmacological modulation consistent with shifts in cortical E/I balance, this study confirms its physiological relevance and supports its use as a biomarker of excitability disturbances and cortical dysfunction across neurological and psychiatric disorders.

Background/Objectives: A sub-anesthetic dose of ketamine has shown promise in reducing craving, withdrawal symptoms, and use of drugs such as alcohol, cocaine, and opioids among individuals with substance use disorders. Ketamine's therapeutic potential for tobacco use is unknown. Here, we investigated a single sub-anesthetic dose among adults with tobacco use disorder who were not interested in changing their smoking behavior. Methods: Utilizing a randomized, within-subject crossover, double-blinded, counter-balanced, midazolam-controlled design, participants (n = 18) received a 0.71 mg/kg infusion of ketamine and a 0.025 mg/kg infusion of midazolam (i.e., active placebo) at least two weeks apart. Participants were asked to abstain from smoking after the infusions until the post-infusion sessions, 1 day following infusion, where participants completed measures of smoking behavior, craving, and withdrawal symptoms. Participants continued to record their smoking behavior over the 7 days following infusion. Participants also completed a semi-structured qualitative interview regarding their experiences. Results: Compared to midazolam, ketamine infusion led to a non-significant reduction (p = 0.10, ηp[2] = 0.153) in the number of cigarettes smoked during the requested abstinence period. Following this period, there were no significant differences in ad lib smoking. Ketamine showed no effect on craving or withdrawal symptoms. Participants reported more intense psychological experiences following ketamine infusion (p < 0.001, ηp[2] = 0.830) and about half reported it felt easier to abstain from smoking after the ketamine infusion. Conclusions: While well tolerated, these findings suggest ketamine has little to no direct effect on quantitative measures of cigarette smoking, craving, or withdrawal. However, the qualitative measures suggest ketamine improves mood and reduces craving in some individuals for several days. Future studies should investigate whether ketamine can indirectly support smoking cessation among individuals with comorbid psychiatric indications for ketamine treatment.

Diuretic resistance in worsening heart failure (HF) is associated with poorer outcomes and, although common, can be challenging to identify. The recently developed BAN-ADHF score can potentially identify patients with worsening HF at risk of diuretic resistance. We externally validated the BAN-ADHF diuretic-resistance score in a multicentre randomized trial comparing adjunctive dapagliflozin to metolazone in patients with HF and diuretic resistance. We calculated a seven-variable modified BAN-ADHF score at baseline and dichotomised patients by the median BAN-ADHF score (8). We assessed loop diuretic efficiency (kg weight change per 40-mg furosemide) and decongestion (weight, eight-zone lung ultrasound B-lines, clinical congestion score) over 96 h. A baseline score ≥8 identified lower loop diuretic efficiency (pooled difference -0.12 kg/40 mg, 95% CI -0.18 to -0.06; p < 0.001) despite similar congestion markers at baseline. The BAN-ADHF score may be useful clinically to identify patients at risk of diminished response to diuretics.

Low energy availability is a core feature of Relative Energy Deficiency in Sport that impairs skeletal and hematologic health. However, the contributions of short-term energy restriction and exercise as well as their potential interactive effects remain unclear. This study aimed to clarify the effects of short-term food restriction on exercise-induced bone mass acquisition and its simultaneous impact on iron metabolism in young female rats. Female Sprague-Dawley rats (5 wk old) underwent environmental acclimation and treadmill running exercise training for 1 wk each and randomly grouped into seven groups. The groups included a sedentary and ad libitum feeding group, along with groups fed 90%, 70%, or 50% of the average ad-lib intake under sedentary or running exercise conditions for 15 d. The results showed low tibial bone mineral density (BMD) and ferritin levels and high serum iron levels due to a significant main effect of food restriction. Hematocrit-adjusted erythrocyte counts were unchanged, indicating an iron metabolic disturbance characterized by depleted stores despite high circulating iron. Exercise significantly increased tibial BMD, but the 50% food restriction groups (sedentary and exercise) showed significantly lower values than the exercise 90% group. Serum iron levels showed significant differences even in the 70% food restriction group compared with the 90% group. Tibial BMD and serum iron levels were weakly negatively correlated. This study revealed that, in young female rats, short-term 50% food restriction resulted in decreased BMD while short-term 70% and 50% food restriction resulted in iron metabolic disturbances under both sedentary and exercise conditions.