An in vivo RNAi screen uncovers the role of AdoR signaling and adenosine deaminase in controlling intestinal stem cell activity.

Metabolites are increasingly appreciated for their roles as signaling molecules. To dissect the roles of metabolites, it is essential to understand their signaling pathways and their enzymatic regulations. From an RNA interference (RNAi) screen for regulators of intestinal stem cell (ISC) activity in the Drosophila midgut, we identified adenosine receptor (AdoR) as a top candidate gene required for ISC proliferation. We demonstrate that Ras/MAPK and Protein Kinase A (PKA) signaling act downstream of AdoR and that Ras/MAPK mediates the major effect of AdoR on ISC proliferation. Extracellular adenosine, the ligand for AdoR, is a small metabolite that can be released by various cell types and degraded in the extracellular space by secreted adenosine deaminase. Interestingly, down-regulation of adenosine deaminase-related growth factor A (Adgf-A) from enterocytes is necessary for extracellular adenosine to activate AdoR and induce ISC overproliferation. As Adgf-A expression and its enzymatic activity decrease following tissue damage, our study provides important insights into how the enzymatic regulation of extracellular adenosine levels under tissue-damage conditions facilitates ISC proliferation.

Euglycemic diabetic ketoacidosis associated with SGLT2i use: Case series.

INTRODUCTION: Euglycemic diabetic ketoacidosis (EuDKA) associated with Sodium-Glucose Cotransporter-2 inhibitor (SGLT2i) use has been described but remains poorly understood. Data on Emergency Department (ED) presentation, resource utilization, and safety outcomes for these patients are lacking. We report a case series of patients diagnosed with EuDKA in the ED. METHODS: An electronic medical record search identified adult patients presenting to a large tertiary ED with EuDKA. They were screened for concurrent use of SGLT2i. Clinical presentation, resource utilization, safety, and disposition data were collected and described. RESULTS: Five patients were included for analysis. Median age [range] was 57 [43-73] years. Presenting symptoms included nausea, vomiting, fatigue, and altered mental status. Initial results included: serum glucose 191 mg/dL [176-215], venous pH 7.01 [6.95-7.30], serum HCO3 8 mEq/L [6-13], anion gap 27 [26-31], serum beta-hydroxybutyrate 9.9 mmol/L [9.2-12.3], and urine ketones 150 [150-150]. Patients remained on an insulin infusion for 18.77 h [11.25-56.48]. There were zero episodes of hypoglycemia and one episode of hypokalemia while on insulin infusion. Time to resolution of metabolic acidosis was 23.82 h [15.45-24.77]. DISCUSSION: We report a case series of ED patients with EuDKA associated with SGLT2i use, and describe presentation characteristics, resource utilization, and safety outcomes. Emergency physicians should be aware of the association between SGLT2i use and EuDKA. An appropriate work-up should be pursued for patients taking an SGLT2i who present with symptoms suggestive of DKA, including nausea, vomiting, malaise, and altered mental status, or are noted to have an unexplained elevated anion gap metabolic acidosis.

The Inpatient Discharge Lounge as a Potential Mechanism to Mitigate Emergency Department Boarding and Crowding.

Delayed access to inpatient beds for admitted patients contributes significantly to emergency department (ED) boarding and crowding, which have been associated with deleterious patient safety effects. To expedite inpatient bed availability, some hospitals have implemented discharge lounges, allowing discharged patients to depart their inpatient rooms while awaiting completion of the discharge process or transportation. This conceptual article synthesizes the evidence related to discharge lounge implementation practices and outcomes. Using a conceptual synthesis approach, we reviewed the medical and gray literature related to discharge lounges by querying PubMed, Google Scholar, and Google and undertaking backward reference searching. We screened for articles either providing detailed accounts of discharge lounge implementations or offering conceptual analysis on the subject. Most of the evidence we identified was in the gray literature, with only 3 peer-reviewed articles focusing on discharge lounge implementations. Articles generally encompassed single-site descriptive case studies or expert opinions. Significant heterogeneity exists in discharge lounge objectives, features, and apparent influence on patient flow. Although common barriers to discharge lounge performance have been documented, including underuse and care team objections, limited generalizable solutions are offered. Overall, discharge lounges are widely endorsed as a mechanism to accelerate access to inpatient beds, yet the limited available evidence indicates wide variation in design and performance. Further rigorous investigation is required to identify the circumstances under which discharge lounges should be deployed, and how discharge lounges should be designed to maximize their effect on hospitalwide patient flow, ED boarding and crowding, and other targeted outcomes.

Effects of residential summer camp on body mass index and body composition in type 1 diabetes.

BACKGROUND: Body mass index (BMI) and fat mass may be higher in children with diabetes compared to healthy peers. It is not certain how diabetic children respond to exercise and diet interventions. OBJECTIVE: To investigate the effect of summer camp on BMI and body composition in children with type 1 diabetes. METHODS: Five hundred eighty-six children (5-19 years, 518 with type 1 diabetes, 68 without diabetes) were followed while attending camp. BMI z-scores (BMIz) and body composition (bioelectrical impedance analysis) were measured at the beginning and end of each 19-day session. Diet and activity were directly supervised, blood glucose closely monitored. A nested diabetic/non-diabetic sib pair analysis was also conducted. Changes in BMIz and percent fat mass (%FM) were the primary outcomes. Findings were confirmed by analysis of data from 612 campers (549 with diabetes) the following summer. RESULTS: At entry, campers with diabetes had higher BMIz and %FM. They tended to gain BMIz (0.04 ± 0.01) whereas non-diabetic campers lost (-0.16 ± 0.11, P < .0001). BMIz increases were positively correlated with precamp hemoglobin A1c values. The differences in initial values and changes in BMIz remained when campers with diabetes were compared to their siblings. All experienced a similar reduction in %FM. Similar results were obtained the following summer. CONCLUSIONS: Children with diabetes may, therefore, accrue more lean body tissue with increased exercise and a healthy diet than those without diabetes. This effect is greatest in those with initially poor metabolic control.

Effect of meal composition on postprandial glucagon-like peptide-1, insulin, glucagon, C-peptide, and glucose responses in overweight/obese subjects.

BACKGROUND: Glucagon-like peptide-1 (GLP-1), an incretin hormone, is released in response to food intake. It is unclear how meals high in protein (HP) and monounsaturated fat (HMF) affect GLP-1 response. PURPOSE: To examine the effect of a HP versus a HMF meal on GLP-1 response. METHODS: Twenty-four overweight/obese participants consumed two meals (HP: 31.9 % energy from protein; HMF: 35.2 % fat and 20.7 % monounsaturated fat) in a random order. Both meals contained the same energy and carbohydrate content. GLP-1, insulin, glucagon, C-peptide, and glucose were assessed from blood drawn in the fasting and postprandial states. The effect of meal condition on hormone and glucose responses and appetite ratings were assessed by repeated measures analysis. RESULTS: Statistically significant (p < 0.01) time by meal condition effect was observed on active GLP-1, total GLP-1, insulin, C-peptide, and glucagon, but not glucose (p = 0.83). Area under the curve was significantly higher during the HP versus the HMF meal conditions for active GLP-1 (23.7 %; p = 0.0007), total GLP-1 (12.2 %; p < 0.0001), insulin (54.4 %; p < 0.0001), C-peptide (14.8 %; p < 0.0001), and glucagon (40.7 %; p < 0.0001). Blood glucose was not different between the HP versus HMF conditions (-4.8 %; p = 0.11). Insulin sensitivity was higher during the HMF versus HP conditions (Matsuda index mean difference: 16.3 %; p = 0.007). Appetite ratings were not different by meal condition. CONCLUSIONS: GLP-1 and insulin responses were higher during the HP condition. However, no difference was found in blood glucose between conditions, and insulin sensitivity was higher during the HMF condition, indicating that a HMF meal may be optimal at regulating blood glucose in overweight/obese individuals without type 2 diabetes.

Temperature Measurement Inside Protective Headgear: Comparison With Core Temperatures and Indicators of Physiological Strain During Exercise in a Hot Environment.

Non-invasive temperature monitoring with a sensor inside protective headgear may be effective in detecting temperatures that are associated with heat illness. The purpose was to establish the relationship between in-hardhat temperatures (Tih) and core temperature (Tc) as measured by rectal (Tre) and esophageal (Tes) probes. Thirty males (age 24.57 ± 4.32 yrs.) completed two trials: continuous submaximal exercise (CSE) and a series of high intensity 30-s sprints (HIE) with a one-minute rest between each. Exercise in both conditions was in a 36(°)C environment (40% RH) while wearing a standard hardhat with sensors mounted on the forehead that were monitored remotely. Exercise continued until voluntary termination or until Tc reached 39.5(°)C. Temperatures, heart rate, cardiorespiratory, and perceptual responses were monitored throughout. A physiological strain index (PSI) was calculated from Tc and HR. The final temperatures in the CSE condition were 38.77 ± 0.41, 38.90 ± 0.49 and 39.29 ± 0.58(°)C and in the HIE condition, final temperatures were 38.76 ± 0.37, 38.91 ± 0.47, and 39.19 ± 0.57 f (o)C for Tih, Tre, and Tes, respectively. The PSI in CSE was 9.62 ± 062, 9.18 ± 1.11, and 10.04 ± 1.05, and in the HIE condition 9.67 ± 068, 9.29 ± 0.99. and 9.86 ± 1.02 based on Tih, Tre and Tes, respectively. The general agreement between the Tih and other temperature measures along with the consistency as indicated by a low coefficient of variation (approx. 1%) in the recordings of the Tih sensors at the point of termination suggest that this device, or similar devices, may have application as a warning system for impending heat-related problems.

Portable Remote Imaging Spectrometer coastal ocean sensor: design, characteristics, and first flight results.

The design, characteristics, and first test flight results are described of the Portable Remote Imaging Spectrometer, an airborne sensor specifically designed to address the challenges of coastal ocean remote sensing. The sensor incorporates several technologies that are demonstrated for the first time, to the best of our knowledge, in a working system in order to achieve a high performance level in terms of uniformity, signal-to-noise ratio, low polarization sensitivity, low stray light, and high spatial resolution. The instrument covers the 350-1050 nm spectral range with a 2.83 nm sampling per pixel, and a 0.88 mrad instantaneous field of view, with 608 cross-track pixels in a pushbroom configuration. Two additional infrared channels (1240 and 1610 nm) are measured by a spot radiometer housed in the same head. The spectrometer design is based on an optically fast (F/1.8) Dyson design form coupled to a wide angle two-mirror telescope in a configuration that minimizes polarization sensitivity without the use of a depolarizer. A grating with minimum polarization sensitivity and broadband efficiency was fabricated as well as a slit assembly with black (etched) silicon surface to minimize backscatter. First flight results over calibration sites as well as Monterey Bay in California have demonstrated good agreement between in situ and remotely sensed data, confirming the potential value of the sensor to the coastal ocean science community.

Natural variation in starvation sensitivity maps to a point mutation in phospholipase IPLA2-VIA in Drosophila melanogaster.

Resistance to starvation is a classic complex trait, where genetic and environmental variables can greatly modify an animal's ability to survive without nutrients. In this study, we investigate the genetic basis of starvation resistance using complementary quantitative and classical genetic mapping in Drosophila melanogaster . Using the Drosophila Genetics Reference Panel (DGRP) as a starting point, we queried the genetic basis of starvation sensitivity in one of the most sensitive DGRP lines. We localize a major effect locus modifying starvation resistance to the phospholipase iPLA2-VIA. This finding is consistent with the work of others which demonstrate the importance of lipid regulation in starvation stress. Furthermore, we demonstrate that iPLA2-VIA plays a role in the maintenance of sugar reserves post-starvation, which highlights a key dynamic between lipid remodeling, sugar metabolism and resistance to starvation stress.

Hospital Capacity Command Centers: A Benchmarking Survey on an Emerging Mechanism to Manage Patient Flow.

BACKGROUND: Delayed hospital and emergency department (ED) patient throughput, which occurs when demand for inpatient care exceeds hospital capacity, is a critical threat to safety, quality, and hospital financial performance. In response, many hospitals are deploying capacity command centers (CCCs), which co-locate key work groups and aggregate real-time data to proactively manage patient flow. Only a narrow body of peer-reviewed articles have characterized CCCs to date. To equip health system leaders with initial insights into this emerging intervention, the authors sought to survey US health systems to benchmark CCC motivations, design, and key performance indicators. METHODS: An online survey on CCC design and performance was administered to members of a hospital capacity management consortium, which included a convenience sample of capacity leaders at US health systems (N = 38). Responses were solicited through a targeted e-mail campaign. Results were summarized using descriptive statistics. RESULTS: The response rate was 81.6% (31/38). Twenty-five respondents were operating CCCs, varying in scope (hospital, region of a health system, or entire health system) and number of beds managed. The most frequent motivation for CCC implementation was reducing ED boarding (n = 24). The most common functions embedded in CCCs were bed management (n = 25) and interhospital transfers (n = 25). Eighteen CCCs (72.0%) tracked financial return on investment (ROI); all reported positive ROI. CONCLUSION: This survey addresses a gap in the literature by providing initial aggregate data for health system leaders to consider, plan, and benchmark CCCs. The researchers identify motivations for, functions in, and key performance indicators used to assess CCCs. Future research priorities are also proposed.

A Generalized Method for Determining Free Soluble Phenolic Acid Composition and Antioxidant Capacity of Cereals and Legumes.

Phenolic acids are a class of organic compounds that bear both a phenolic group, and a carboxylic group. They are found in grains and concentrate in the bran of cereals or seed coat of legumes. They possess antioxidant properties that have generated much research interest in recent years, about their potential antioxidant protective health functions. This work presents a generalized method for the extraction of free soluble phenolic acids from whole grains and analysis of their antioxidant capacity. Five whole grain samples comprising two cereals (wheat and yellow corn) and three legumes (cowpea bean, kidney bean, and soybean), were used. The grains were milled into flour and their free soluble phenolic acids extracted using aqueous methanol. The compounds were then identified using a high-pressure liquid chromatograph (HPLC). The Folin-Ciocalteu method was used to determine their total phenolic content while their antioxidant capacities were determined using the DPPH radical scavenging capacity, Trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. The phenolic acids identified included vanillic, caffeic, p-coumaric and ferulic acids. Vanillic acid was identified only in cowpea while caffeic acid was identified only in kidney bean. p-Coumaric acid was identified in yellow corn, cowpea, and soybean, while ferulic acid was identified in all the samples. Ferulic acid was the predominant phenolic acid identified. The total concentration of phenolic acids in the samples decreased in the following order: soybean > cowpea bean > yellow corn = kidney bean > wheat. The total antioxidant capacity (sum of values of DPPH, TEAC and ORAC assays) decreased as follows: soybean > kidney bean > yellow corn = cowpea bean > wheat. This study concluded that HPLC analysis as well as DPPH, TEAC, and ORAC assays provide useful information about the phenolic acid composition and antioxidant properties of whole grains.

Use of Hospital Capacity Command Centers to Improve Patient Flow and Safety: A Scoping Review.

OBJECTIVES: Delayed emergency department (ED) and hospital patient throughput is recognized as a critical threat to patient safety. Increasingly, hospitals are investing significantly in deploying command centers, long used in airlines and the military, to proactively manage hospital-wide patient flow. This scoping review characterizes the evidence related to hospital capacity command centers (CCCs) and synthesizes current data regarding their implementation. METHODS: As no consensus definition exists for CCCs, we characterized them as units (i) involving interdisciplinary, permanently colocated teams, (ii) using real-time data, and (iii) managing 2 or more patient flow functions (e.g., bed management, transfers, discharge planning, etc.), to distinguish CCCs from transfer centers. We undertook a scoping review of the medical and gray literature published through April 2019 related to CCCs meeting these criteria. RESULTS: We identified 8 eligible articles (including 4 peer-reviewed studies) describing 7 CCCs of varying designs. The most common CCC outcome measures related to transfer volume (n = 5) and ED boarding (n = 4). Several CCCs also monitored patient-level clinical parameters. Although all articles reported performance improvements, heterogeneity in CCC design and evidence quality currently restricts generalizability of findings. CONCLUSIONS: Numerous anecdotal accounts suggest that CCCs are being widely deployed in an effort to improve hospital patient flow and safety, yet peer-reviewed evidence regarding their design and effectiveness is in its earliest stages. The costs, objectives, and growing deployment of CCCs merit an investment in rigorous research to better measure their processes and outcomes. We propose a standard definition, conceptual framework, research priorities, and reporting standards to guide future investigation of CCCs.

Phenolic Composition and Antioxidant Properties of Cooked Rice Dyed with Sorghum-Leaf Bio-Colorants.

White rice is an important staple food globally. It is a rich source of energy but is low in dietary phenolic antioxidants. This current research aimed at providing scientific evidence for an alternative rice dish that has increased phenolic-antioxidant health-promoting potential by combining white rice with red cowpea beans and cooking with dye sorghum leaves hydrothermal extract, as a source of natural colorant. Boiled white rice and the rice-cowpea-sorghum extract dish were freeze-dried, and the free and bound phenolic compounds of raw and cooked samples were extracted. Phenolic composition, total phenolic content (TPC), and antioxidant activities (measured by 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity, Trolox equivalent antioxidant capacity, and oxygen radical absorbance capacity methods) of the raw and cooked samples were determined. Combining white rice with cowpea seeds and sorghum leaves extract significantly (p < 0.0001) increased the TPC and antioxidant activities of the rice due to the higher TPC and antioxidant activities of cowpea and sorghum leaves. Although boiling caused substantial losses of flavonoids and anthocyanins in the rice-cowpea-sorghum extract composite meal, the resulting dish had higher TPC and antioxidant activities than boiled white rice. Compositing white rice with phenolic-rich pulses can be an innovative approach to providing alternative healthy rice dishes to consumers.

Emergency department provider in triage: assessing site-specific rationale, operational feasibility, and financial impact.

Emergency department (ED) crowding is recognized as a critical threat to patient safety, while sub-optimal ED patient flow also contributes to reduced patient satisfaction and efficiency of care. Provider in triage (PIT) programs-which typically involve, at a minimum, a physician or advanced practice provider conducting an initial screening exam and potentially initiating treatment and diagnostic testing at the time of triage-are frequently endorsed as a mechanism to reduce ED length of stay (LOS) and therefore mitigate crowding, improve patient satisfaction, and improve ED operational and financial performance. However, the peer-reviewed evidence regarding the impact of PIT programs on measures including ED LOS, wait times, and costs (as variously defined) is mixed. Mechanistically, PIT programs exert their effects by initiating diagnostic work-ups earlier and, sometimes, by equipping triage providers to directly disposition patients. However, depending on local contextual factors-including the co-existence of other front-end interventions and delays in ED throughput not addressed by PIT-we demonstrate how these features may or may not ultimately translate into reduced ED LOS in different settings. Consequently, site-specific analysis of the root causes of excessive ED LOS, along with mechanistic assessment of potential countermeasures, is essential for appropriate deployment and successful design of PIT programs at individual EDs. Additional motivations for implementing PIT programs may include their potential to enhance patient safety, patient satisfaction, and team dynamics. In this conceptual article, we address a gap in the literature by demonstrating the mechanisms underlying PIT program results and providing a framework for ED decision-makers to assess the local rationale for, operational feasibility of, and financial impact of PIT programs.

Impact of multidisciplinary team huddles on patient safety: a systematic review and proposed taxonomy.

BACKGROUND: Despite significant advances, patient safety remains a critical public health concern. Daily huddles-discussions to identify and respond to safety risks-have been credited with enhancing safety culture in operationally complex industries including aviation and nuclear power. More recently, huddles have been endorsed as a mechanism to improve patient safety in healthcare. This review synthesises the literature related to the impact of hospital-based safety huddles. METHODS: We conducted a systematic review of peer-reviewed literature related to scheduled, multidisciplinary, hospital-based safety huddles through December 2019. We screened for studies (1) in which huddles were the primary intervention being assessed and (2) that measured the huddle programme's apparent impact using at least one quantitative metric. RESULTS: We identified 1034 articles; 24 met our criteria for review, of which 19 reflected unit-based huddles and 5 reflected hospital-wide or multiunit huddles. Of the 24 included articles, uncontrolled pre-post comparison was the prevailing study design; we identified only two controlled studies. Among the 12 unit-based studies that provided complete measures of statistical significance for reported outcomes, 11 reported statistically significant improvement among some or all outcomes. The objectives of huddle programmes and the language used to describe them varied widely across the studies we reviewed. CONCLUSION: While anecdotal accounts of successful huddle programmes abound and the evidence we reviewed appears favourable overall, high-quality peer-reviewed evidence regarding the effectiveness of hospital-based safety huddles, particularly at the hospital-wide level, is in its earliest stages. Additional rigorous research-especially focused on huddle programme design and implementation fidelity-would enhance the collective understanding of how huddles impact patient safety and other targeted outcomes. We propose a taxonomy and standardised reporting measures for future huddle-related studies to enhance comparability and evidence quality.

Carica papaya seed enhances phytochemicals and functional properties in cornmeal porridges.

Papaya seeds, a high source of dietary nutrients and phytochemicals are wasted when Carica papaya fruit is processed and consumed. This study investigates bioactivity of papaya seeds (PS) from 3 different locations in Kenya for potential valorization as porridge. PS was treated with acetic acid and sodium bicarbonate to improve pallatability. HPLC analysis revealed that PS flour added compounds which were absent from cornmeal (p-hydroxybenzoic, 2,4-dihydroxybenzoic and vanillic acids) and increased over 25% the pre-existing ones. Acid and alkali treatments increased the phenolic compounds content and antioxidant capacities of the seed 1 porridges in ≈19% average. The differential scanning calorimetry and the rapid visco analysis showed a significant decrease in the enthalpy required (≈44%) to gelatinize cornmeal-PS blend and the tendency for retrogradation (from 2188 to 700 cP average). Therefore, our findings indicate that PS can contribute to improved phytochemical and functional properties of cornmeal porridges.

A cytosine deaminase for programmable single-base RNA editing.

Programmable RNA editing enables reversible recoding of RNA information for research and disease treatment. Previously, we developed a programmable adenosine-to-inosine (A-to-I) RNA editing approach by fusing catalytically inactivate RNA-targeting CRISPR-Cas13 (dCas13) with the adenine deaminase domain of ADAR2. Here, we report a cytidine-to-uridine (C-to-U) RNA editor, referred to as RNA Editing for Specific C-to-U Exchange (RESCUE), by directly evolving ADAR2 into a cytidine deaminase. RESCUE doubles the number of mutations targetable by RNA editing and enables modulation of phosphosignaling-relevant residues. We apply RESCUE to drive ß-catenin activation and cellular growth. Furthermore, RESCUE retains A-to-I editing activity, enabling multiplexed C-to-U and A-to-I editing through the use of tailored guide RNAs.

Bioactive compounds and biological properties of Brazilian stingless bee honey have a strong relationship with the pollen floral origin.

Multivariate data analysis feasibility for the evaluation of Brazilian stingless bee honey (SBH) by pollen spectrum, bioactive compounds content, physicochemical, antioxidant and antimicrobial analysis was investigated. Levels of total and individual phenolics content were analyzed by HPLC-PDA. The antioxidant capacity was performed by 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH), oxygen radical absorption capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays. The total phenolic compounds from the thirty-two SBH was positively correlated with the antioxidant capacity. Bioactive compounds such as p-coumaric acid, quercetin, and hesperetin were identified in all the samples. Brazilian SBH shows more effective antibacterial activity against Gram-negative bacteria (E. coli and S. Typhimurium) compared to Gram-positive ones. Results also revealed that SBH could reach up to 45% higher antioxidant and biological activities than the traditional Apis mellifera honey. Chemometrics shows that chemical and biological properties of SBH have a strong relationship with the pollen botanical origin. Principal component analysis (PCA) grouped the honey into three categories with predominant pollen from Verbenaceae, Asteraceae and Sapindaceae families, confirming that SBH belonging to the same floral origin present similar characteristics.

The Effects of High-Protein and High-Monounsaturated Fat Meals on Postprandial Lipids, Lipoprotein Particle Numbers, Cytokines, and Leptin Responses in Overweight/Obese Subjects.

BACKGROUND: Obesity is linked to dyslipidemia, proinflammatory state, and hyperleptinemia. The influence of high-protein (HP) versus high-monounsaturated fat (HMF) meals on postprandial lipids, lipoprotein particle numbers, cytokines, and leptin responses in overweight/obese (OW/O) subjects is unknown. METHODS: Twenty-four OW/O participants consumed an HP (31.9% energy from protein) and HMF (35.2% fat and 20.7% monounsaturated fat) meal, of similar energy/carbohydrate content, in a random order. The outcome variables were assessed from blood samples collected in fasted and postprandial (3 hr) states. RESULTS: Repeated measures analysis found significant (P < 0.05) meal condition by time interactions for triglycerides (TGs), very low-density lipoprotein particles (VLDLP), total high-density lipoprotein particles (T-HDLP), and the ratio of large-buoyant high-density lipoprotein 2b (LB-HDL2b) to T-HDLP, and meal effect on small-dense HDLP (SD-HDLP). Comparison of HP versus HMF condition showed significantly lower TG at 120 min [geometric mean (95% confidence interval, CI): 148 (125-175) vs. 194 (164-230) mg/dL] and 180 min [167 (138-203) vs. 230 (189-278) mg/dL] and VLDLP at 180 min [70.0 (58.2-84.3) vs. 88.0 (73.1-106) nmol/L]. HP versus HMF condition showed significantly lower LB-HDL2b/T-HDLP at 180 min [mean difference (95% CI): 0.021 (0.004-0.038)], and higher T-HDLP [671 (263-1079) nmol/L] and SD-HDLP [606 (292-920) nmol/L] at 120 min. Area under the curve was significantly lower for TG and higher for T-HDLP, SD-HDLP, and small-dense LDL III (SD-LDL III) in the HP condition. Cytokines and leptin were not different between conditions. CONCLUSION: OW/O subjects had lower TG and VLDLP, but less favorable SD-LDL III, SD-HDLP, and LB-HDL2b/T-HDLP ratio responses to the HP versus HMF meals.

Effect of meal composition on postprandial lipid concentrations and lipoprotein particle numbers: A randomized cross-over study.

BACKGROUND: It is unclear how high-protein (HP) and high-monounsaturated fat (HMF) meals affect postprandial blood lipids and lipoprotein particle numbers (LPN). PURPOSE: To compare a HP versus a HMF meal on postprandial lipid and LPN responses. METHODS: Twenty-four participants (age: 36.3±15.0 years; body mass index: 23.6±2.0 kg/m2; 45.8% female) were fed a HP (31.9% energy from protein) and a HMF (35.2% fat and 20.7% monounsaturated fat) meal in a randomized cross-over trial design. Energy and carbohydrate content were the same across meals. Blood samples were drawn in the fasting state and 3 hour postprandial state, and assessed for lipids and LPN. RESULTS: Repeated measures analysis showed a significant (p<0.05) treatment by time interaction effect for triglycerides (TG), the primary variable, total high-density lipoprotein particles (T-HDLP) and T-HDLP minus large-buoyant high-density lipoprotein 2b (T-HDLP-LB-HDL2b). HP versus HMF condition led to significantly lower TG at 120 (geometric mean: 90.1 (95% confidence interval (CI): 76.4-106.3) vs. 146.5 (124.2-172.9) mg/dL) and 180 (101.4 (83.1-123.8) vs. 148.7 (121.9-181.4) mg/dL) min and higher T-HDLP at 120 (mean difference: 297.3 (95% CI: 48.6-545.9) nmol/L) and 180 (291.6 (15.8-567.5) nmol/L) min. The difference in T-HDLP by condition was due to the significantly higher small-dense HDLP (T-HDLP-LB-HDL2b) during HP versus HMF condition at 120 (mean difference: 452.6 (95% CI: 177.4-727.9) nmol/L) and 180 (496.8 (263.1-730.6) nmol/L) min. Area under the curve analysis showed that HP versus HMF condition led to significantly lower TG, non-HDLP, and very-low-density lipoprotein particles (VLDLP) responses but significantly less favorable responses in LB-HDL2b particles, T-HDLP-LB-HDL2b, and LB-HDL2b/T-HDLP ratio. CONCLUSION: The HP meal led to lower TG, non-HDLP, and VLDLP but less favorable LB-HDL2b, small-dense HDLP, and LB-HDL2b/T-HDLP ratio responses versus a HMF meal. Further studies are needed to confirm these findings over multiple meals.

RNA editing with CRISPR-Cas13.

Nucleic acid editing holds promise for treating genetic disease, particularly at the RNA level, where disease-relevant sequences can be rescued to yield functional protein products. Type VI CRISPR-Cas systems contain the programmable single-effector RNA-guided ribonuclease Cas13. We profiled type VI systems in order to engineer a Cas13 ortholog capable of robust knockdown and demonstrated RNA editing by using catalytically inactive Cas13 (dCas13) to direct adenosine-to-inosine deaminase activity by ADAR2 (adenosine deaminase acting on RNA type 2) to transcripts in mammalian cells. This system, referred to as RNA Editing for Programmable A to I Replacement (REPAIR), which has no strict sequence constraints, can be used to edit full-length transcripts containing pathogenic mutations. We further engineered this system to create a high-specificity variant and minimized the system to facilitate viral delivery. REPAIR presents a promising RNA-editing platform with broad applicability for research, therapeutics, and biotechnology.