Sequence-Controlled Spherical Nucleic Acids: Gene Silencing, Encapsulation, and Cellular Uptake.

Antisense oligonucleotides (ASOs) can predictably alter RNA processing and control protein expression; however, challenges in the delivery of these therapeutics to specific tissues, poor cellular uptake, and endosomal escape have impeded progress in translating these agents into the clinic. Spherical nucleic acids (SNAs) are nanoparticles with a DNA external shell and a hydrophobic core that arise from the self-assembly of ASO strands conjugated to hydrophobic polymers. SNAs have recently shown significant promise as vehicles for improving the efficacy of ASO cellular uptake and gene silencing. However, to date, no studies have investigated the effect of the hydrophobic polymer sequence on the biological properties of SNAs. In this study, we created a library of ASO conjugates by covalently attaching polymers with linear or branched [dodecanediol phosphate] units and systematically varying polymer sequence and composition. We show that these parameters can significantly impact encapsulation efficiency, gene silencing activity, SNA stability, and cellular uptake, thus outlining optimized polymer architectures for gene silencing.

Correlation between ketones and mental fatigue in high fat-induced obese and non-obese rats.

Obesity, often caused by a diet high in calories and low physical activity, may induce physical fatigue, as experienced via decreased locomotor activity and mental fatigue such as impaired cognition. This study aims to evaluate glucose and ketone levels secondary to high-fat diet (HFD) exposure and signs of physical and mental fatigue. Fifty-four 7-week-old male Sprague Dawley rats (Rattus norvegicus) were assigned to either an HFD (n = 28) or a standard diet (SD; n = 26) for a 6-week period during which body weight, blood glucose, and ketones were measured twice per week. An open field (OF) paradigm was used to measure locomotor activity, while novel object recognition (NOR) test was used as an indicator of cognition. Animals in the HFD group weighed more than SD rats (8.4 g; p < 0.05) starting at Day 11, blood glucose levels were higher in the HFD group versus SD rats (3.9 mg/dl; p < 0.05) beginning in Week 5, and ketones were lower for the HFD versus the SD group throughout the study (0.34 mmol/L on average; p < 0.05). Although there was no significant difference in locomotor activity between the HFD and SD groups (p = 0.12), regardless of diet, higher ketone levels were associated with increased NOR time and ratio between the familiar and novel objects (p < 0.01). Thus, this study provides evidence that an increased level of ketones is associated with greater cognitive performance and a lesser probability of experiencing mental fatigue.

Design and enhanced gene silencing activity of spherical 2'-fluoroarabinose nucleic acids (FANA-SNAs).

Drug delivery vectors for nucleic acid therapeutics (NATs) face significant barriers for translation into the clinic. Spherical nucleic acids (SNAs) - nanoparticles with an exterior shell made up of DNA strands and a hydrophobic interior - have recently shown great potential as vehicles to improve the biodistribution and efficacy of NATs. To date, SNA design has not taken advantage of the powerful chemical modifications available to NATs. Here, we modify SNAs with 2'-deoxy-2'-fluoro-d-arabinonucleic acid (FANA-SNA), and show increased stability, enhanced gene silencing potency and unaided uptake (gymnosis) as compared to free FANA. By varying the spacer region between the nucleic acid strand and the attached hydrophobic polymer, we show that a cleavable DNA based spacer is essential for maximum activity. This design feature will be important when implementing functionalized nucleic acids into nanostructures for gene silencing. The modularity of the FANA-SNA was demonstrated by silencing two different targets. Transfection-free delivery was superior for the modified SNA compared to the free FANA oligonucleotide.

Daily Self-Weighing to Prevent Holiday-Associated Weight Gain in Adults.

OBJECTIVE: Holiday weight gain is reported to be 0.4 to 1.5 kg and may contribute to annual weight gain. The objective of this study was to test the efficacy of daily self-weighing (DSW) using visual graphical feedback (GF) to prevent holiday weight gain. METHODS: A total of 111 adults were randomly assigned into a control or DSW + GF group and completed the preholiday visit (v1; before Thanksgiving), the postholiday visit (v2; after New Year's Day), and the follow-up visit (v3; 14 weeks after v2). The participants in the DSW + GF group performed DSW with Wi-Fi scales during the holidays and were told to try not to gain weight above baseline weight. RESULTS: There was no change in weight with DSW + GF, whereas the control group gained weight from v1 to v2 (-0.13 ± 0.27 kg vs. 2.65 ± 0.33 kg, P < 0.001, respectively). In the control group, weight change was similar between individuals with overweight or obesity (OW/OB) versus individuals with normal weight (2.71 ± 0.48 kg vs. 2.62 ± 0.43 kg, not significant, respectively). For DSW + GF, individuals with OW/OB lost weight whereas those with normal weight maintained weight during the holidays (-1.46 ± 0.62 kg vs. 0.33 ± 0.27 kg, P = 0.01, respectively). The control group lost weight during the follow-up (-1.14 ± 0.43 kg, P = 0.01; v2 to v3) but retained 57% of weight gain; therefore, weight gain from v1 to v3 was significant (1.51 ± 0.39 kg, P < 0.001). CONCLUSIONS: DSW + GF was a successful approach to prevent holiday weight gain, with those with OW/OB responding most favorably to DSW + GF.

A 7-day high-PUFA diet reduces angiopoietin-like protein 3 and 8 responses and postprandial triglyceride levels in healthy females but not males: a randomized control trial.

BACKGROUND: Polyunsaturated fatty acids (PUFAs) have beneficial effects on hypertriglyceridemia although their effect on angiopoietin-like proteins (ANGPTLs), specifically ANGPTL3, ANGPTL4 and ANGPTL8 is unknown. OBJECTIVE: To determine whether a high-PUFA diet improves postprandial triglyceride (TG) levels through reducing ANGPTL responses following high saturated fat (SFA) meals. METHODS: Twenty-six adults were randomized into a PUFA diet (n = 16) or a control diet group (n = 10). Participants completed a pre-diet visit (v1) where they were given two SFA-rich, high-fat meals. Blood draws were taken at fasting and every 2 h postprandially for a total of 8 h. After v1, participants completed a 7d diet of the same macronutrient proportions (50% carbohydrate, 35% fat, 15% protein) but with different fatty acid (FA) compositions (PUFA = 21% of total energy from PUFAs vs. Control = 7% of total energy from PUFA). All participants then completed the post-diet visit (v2) identical to v1. RESULTS: In the PUFA group, females, but not males, reduced TG concentrations (Area under the curve (AUC): 141.2 ± 18.7 vs. 80.7 ± 6.5 mg/dL/h, p = 0.01, for v1 vs. v2, respectively). Fasting and postprandial AUC levels of ANGPTL3 and 8, but not ANGPTL4, also decreased from v1 to v2 in PUFA females, but not males. No changes from v1 to v2 were seen in either sex in the control group. CONCLUSIONS: A PUFA-rich diet improves TG levels in response to high-SFA meals with reductions in ANGPTL3 and ANGPTL8. PUFAs may be more protective against hypertriglyceridemia in females, compared to males since no diet effect was observed in males. TRIAL REGISTRATION: NCT02246933.

Determining the Accuracy and Reliability of Indirect Calorimeters Utilizing the Methanol Combustion Technique.

BACKGROUND: Several indirect calorimetry (IC) instruments are commercially available, but comparative validity and reliability data are lacking. Existing data are limited by inconsistencies in protocols, subject characteristics, or single-instrument validation comparisons. The aim of this study was to compare accuracy and reliability of metabolic carts using methanol combustion as the cross-laboratory criterion. METHODS: Eight 20-minute methanol burn trials were completed on 12 metabolic carts. Respiratory exchange ratio (RER) and percent O2 and CO2 recovery were calculated. RESULTS: For accuracy, 1 Omnical, Cosmed Quark CPET (Cosmed), and both Parvos (Parvo Medics trueOne 2400) measured all 3 variables within 2% of the true value; both DeltaTracs and the Vmax Encore System (Vmax) showed similar accuracy in measuring 1 or 2, but not all, variables. For reliability, 8 instruments were shown to be reliable, with the 2 Omnicals ranking best (coefficient of variation [CV] < 1.26%). Both Cosmeds, Parvos, DeltaTracs, 1 Jaeger Oxycon Pro (Oxycon), Max-II Metabolic Systems (Max-II), and Vmax were reliable for at least 1 variable (CV ≤ 3%). For multiple regression, humidity and amount of combusted methanol were significant predictors of RER (R2 = 0.33, P < .001). Temperature and amount of burned methanol were significant predictors of O2 recovery (R2 = 0.18, P < .001); only humidity was a predictor for CO2 recovery (R2 = 0.15, P < .001). CONCLUSIONS: Omnical, Parvo, Cosmed, and DeltaTrac had greater accuracy and reliability. The small number of instruments tested and expected differences in gas calibration variability limits the generalizability of conclusions. Finally, humidity and temperature could be modified in the laboratory to optimize IC conditions.