Ad Lib Research Articles

0055 Effect of acute sleep restriction on sleep physiology and emotional imagery memory in schizophrenia

Abstract Introduction Cognitive dysfunction in individuals with schizophrenia (SZ) represents a major determinant of poor functioning and may arise in part due to deficits in sleep quality that have been reported in in this population. In the current project, we investigate the effect of acute sleep restriction on a sleep-dependent emotional memory task. Methods Six individuals with medication-controlled SZ encoded consensus-rated emotionally valent and neutral pictures from the International Affective Picture System in the evening before 2 counterbalanced, polysomnographically-recorded nights containing either ad libitum sleep (~11pm to 7am) or sleep restriction (SR) to 4 hours between 3 and 7 am. Image recall occurred within 1 hour of wake time. We compared memory performance for negative emotional/neutral memory and PSG measures across the two sleep conditions with paired signed-rank tests. Results Mean age was 38 years and 50% were women. Acute sleep restriction resulted in reduced total sleep time (Adlib: median 372 min. (interquartile-range 53) vs SR: 209 (16) min., p=0.03), including reduction in duration of N1 (Ad lib: 44 min. (44) vs. SR: 29 min. (33), p=0.03), N2 (Ad lib: 158 min. (86) vs. SR: 60 min. (33), p=0.03) and REM sleep (Ad lib: 18 min. (5) vs. SR: 13 min. (13), p=0.03). We did not observe differences in N3 duration, sleep efficiency, and arousal index. Measures of breathing such as AHI4%, AHI3A, and SpO2 nadir were not significantly different between conditions. Compared to SR, the false alarm rate for emotional images was increased in ad lib sleep (n=6, p=0.03) with no difference in false alarm rate for neutral images (p=0.19). Correct rejection rate for emotional images was increased in SR compared to ad lib sleep (p=0.03) with no difference in neutral images (p=0.19). No other change was observed in accuracy of recall for emotional or neutral, miss rate for emotional or neutral and d-prime for emotional or neutral images. Conclusion In subjects with SZ, unrestricted sleep was associated with a greater false alarm rate for emotional, but not neutral scenes, suggesting a deficit in veridical memory or generalization of emotionally encoded information. Support (if any) R21MH126357, R01AG066870, R01AG056682, K24HL109156, AASMF Focus award/AARG-D

Variation in the Early Life and Adult Intestinal Microbiome of Intra-Uterine Growth Restricted Rat Offspring Exposed to a High Fat and Fructose Diet.

Intra-Uterine Growth Restriction (IUGR) is a risk factor for many adult-onset chronic diseases, such as diabetes and obesity. These diseases are associated with intestinal microbiome perturbations (dysbiosis). The establishment of an intestinal microbiome begins in utero and continues postnatally (PN). Hypercaloric diet-induced dysbiosis is a major driver of childhood obesity. We hypothesized that different postnatal diets superimposed on IUGR will alter the postnatal intestinal microbiome. We compared four experimental rat groups: (1) Ad lib fed regular chow diet pre- and postnatally (CON), (2-3) IUGR induced by maternal caloric restriction prenatally followed postnatally (PN) by either (2) the control diet (IUGR-RC) or (3) High-Fat-high-fructose (IUGR-HFhf) diet, and lastly (4) HFhf ad lib pre- and postnatally (HFhf). Fecal samples were collected from dams and male and female rat offspring at postnatal day 2, 21, and adult day 180 for 16S rRNA gene sequencing. Maternal diet induced IUGR led to dysbiosis of the intestinal microbiome at PN21. Postnatal HFhf diet significantly reduced microbial diversity and worsened dysbiosis reflected by an increased Gammaproteobacteria/Clostridia ratio. Dysbiosis arising from a mismatch between IUGR and a postnatal HFhf diet may contribute to increased risk of the IUGR offspring for subsequent detrimental health problems.

Acquisition and persistence of intermittent access-induced escalated sucrose intake in male rats

Ad lib fed male rats with 23 h every third day access (E3DA) to 4% sucrose come to double the sucrose intake of rats with everyday access (EDA). These differences are maintained if all rats are then put on an alternate day schedule. In two experiments, we explored how initial patterns of sucrose availability influenced consumption when access conditions changed. In Phase I of a 2 by 2 experiment 1, rats given E3DA to 4% sucrose approximately doubled the intake of rats given EDA. In Phase II, half the EDA rats were shifted to E3DA and they rapidly increased their sucrose intake, but their sucrose consumption remained below that of the Phase I E3DA rats. Rats shifted from E3DA to EDA only gradually reduced their intake over the 40 days. In Phase III, all rats were shifted to alternate day access and the effects of prior phases were still evident. In experiment 2, rats given 40 days of EDA to 4% sucrose and then shifted to E3DA showed almost identical sucrose intake increases as age-matched rats with no previous sucrose experience. The period of EDA to sucrose did not influence the magnitude of the E3DA effect. In a third experiment, we found that older rats showed an intermittency effect similar to what we have found with younger rats. Limited or unpredictable availability of a food source, even after a period of abundance, can produce a profound positive change in an animal's evaluation of the item.

The Study of Australian Jazz and the Issue of Methodological Nationalism

The Study of Australian Jazz and the Issue of Methodological Nationalism

Fat Stores and Antioxidant Capacity Affect Stopover Decisions in Three of Four Species of Migratory Passerines With Different Migration Strategies: An Experimental Approach

During migratory stopovers, birds must make decisions about when and where to travel and these decisions are likely contingent on their fuel stores, food availability, and antioxidant capacity as well as seasonal changes in key environmental factors. We conducted a field experiment on an offshore stopover site (Block Island, Rhode Island, United States: 41°130N, 71°330W) during autumn migration to test the hypothesis that birds with greater fuel stores and non-enzymatic antioxidant capacity have shorter stopovers than lean birds with low antioxidant capacity, and to determine the extent to which this depends on migration strategy. We used a 2 × 2 factorial field experiment (two levels each of available food and dietary polyphenols) with four species of songbirds kept in captivity for 3–5 days to produce experimental groups with different fuel stores and antioxidant capacity. We attached digital VHF transmitters to assess stopover duration and departure direction using automated telemetry. Non-enzymatic antioxidant capacity increased during refueling for Red-eyed Vireos (Vireo olivaceus) and Blackpoll Warblers (Setophaga striata) fedad libdiets, and forad libfed Hermit Thrushes (Catharus guttatus) supplemented with polyphenols, but not for Yellow-rumped Warblers (Setophaga coronata coronata). Glutathione peroxidase (GPx) decreased during captivity and was influenced by dietary treatment only in Red-eyed Vireos. Oxidative damage decreased during captivity for all species except Yellow-rumped Warblers. Stopover duration was shorter for Vireos and Blackpolls fedad libas compared to those fed maintenance.Ad libfed Hermit Thrushes supplemented with polyphenols had shorter stopovers than those fedad lib, as did thrushes fed at maintenance and supplemented with polyphenols compared with those fed at maintenance alone. There was no influence of condition on stopover duration for Yellow-rumped Warblers. Departure direction was not strongly related to condition, and birds primarily reoriented north when departing Block Island. Thus, fat stores and oxidative status interacted to influence the time passerines spent on stopover, and condition-dependent departure decisions were related to a bird’s migration strategy. Therefore, seasonal variation in macro- and micro-nutrient resources available for refueling at stopover sites can affect body condition and antioxidant capacity and in turn influence the timing and success of migration.

Predefined Clinical and Laboratory Criteria Predict Successful Transition from Peanut Oral Immunotherapy Daily Maintenance Dosing to Ad Lib Consumption

Peanut oral immunotherapy has been shown to effectively decrease sensitization in peanut-allergic individuals; however, clinical endpoints for completion of therapy remain unclear. We followed long-term outcomes in a real-world peanut OIT cohort that transitioned to ad lib peanut consumption once meeting predefined clinical and laboratory criteria. Data were collected for 199 patients initiating peanut oral immunotherapy (OIT) at a pediatric Allergy clinic between 2011-2020. Patients proceeded through clinic updosing to daily maintenance dose of 2 teaspoons peanut butter, or lower dose for toddlers or those with severe peanut aversion. Patients were permitted to transition to ad lib peanut consumption once meeting the following criteria: (1) tolerance of a full-serving of peanut (7000mg peanut protein) in oral food challenge, (2) Skin prick wheal <8mm, and (3) serum peanut-specific IgE level <1 kUA/L. Patients were advised to continue peanut consumption at least twice weekly during ad lib regimen. Clinical outcomes were monitored by clinic visits or telephone contact. Fifty-one of 199 patients transitioned to ad lib consumption after a median of 285 days since reaching daily maintenance (91-1183 days). Median time on ad lib status was 581 days (20-2,275 days). Two grade I reactions--oral itching and perioral rash--occurred after lapse in peanut intake for 2 weeks and 2 months, respectively. Eight patients (16%) continued to self-report peanut allergy, and 24 (48%) continued to carry epinephrine injectors. Peanut OIT patients identified prospectively by clinical and laboratory criteria can successfully transition from daily maintenance dosing to intermittent ad lib peanut consumption.

4447 Leptin supplementation prevents the loss of hypoglycemia-induced glucagon release following exposure to six days of severe caloric restriction in mice

OBJECTIVES/GOALS: We have recently shown that mice exposed to six days of 60% caloric restriction acutely display reduced hypoglycemia-induced glucagon release following refeeding, and that this effect is concurrent with low leptin levels. The current study was conducted to ascertain if leptin treatment during caloric restriction would reverse this effect. METHODS/STUDY POPULATION: Three groups of mice were used, an ad libitum (Ad-lib) fed group and two caloric restriction (CR) groups, one of which received twice daily leptin injection (0.5-1μg/g; IP) and the other vehicle (saline) during their caloric restriction. CR mice were placed on 60% caloric restriction for 6 consecutive days. Ad lib mice were housed in an identical manner but fed ad libitum during this same period. Following 6 days of restriction, CR mice were given ad lib access to food for 16 h. After the 16 h period of refeeding, both CR and ad lib mice began a 6 h fast which was immediately followed by a hypoglycemic insulin tolerance test (ITT). ITTs consisted of a variable dose of insulin intended to achieve a blood glucose of ~45 mg/dL within 60 minutes, at which time blood was collected for glucagon and corticosterone assays. RESULTS/ANTICIPATED RESULTS: The mean blood glucose levels during the ITT at 45 and 60 minutes post injection across all three groups were 46.8 + 3.1 and 37.0 + 2.4, respectively. There were no significant differences in glucose levels between the three groups at these two time points. As expected, saline treated CR mice displayed significantly reduced serum glucagon levels in response to the ITT relative to Ad-lib mice (23.5 + 10.9 vs. 91.7 + 20.8 pg/mL, p = 0.009). In contrast, leptin-treated CR mice maintained their hypoglycemia-induced glucagon response to the ITT (78.0 + 16.8 pg/mL, p&gt;0.99 vs. Ad-lib group). In addition, although corticosterone levels in saline treated CR mice were numerically lower than in Ad-lib mice, this difference was not statistically significance (3928 + 277 vs. 4571 + 178 pg/mL, p = 0.179). DISCUSSION/SIGNIFICANCE OF IMPACT: Diabetes patients on insulin therapy often develop impaired hypoglycemic counter-regulation which can lead to life-threatening hypoglycemic complications. Our results suggest that leptin may hold promise as a therapeutic intervention for the prevention of impaired hypoglycemic counter-regulation in persons with diabetes.

PSII-34 Comparison of the activity level of high and low residual feed intake beef steers

Abstract Activity level may affect residual feed intake (RFI), and cattle that move more may have less energy for gain. We compared activity level in high and low RFI beef steers under different nutritional conditions. Lying time, lying duration and lying bouts were recorded at 1-minute intervals utilizing Hobo G Pendant Loggers attached to the right hind leg. To identify RFI classification, 57 Angus-cross steers were individually housed and offered ad libitum access to a TMR for 56 d. RFI was defined as the residual of the regression of DMI on mid-test BW0.75 and ADG. High and low RFI groups (14 steers each) were defined as &amp;gt; 0.5 SD above or below the mean of zero, respectively. Ad libitum feeding was followed by restricted feeding at 75% of previous ad libitum DMI/BW0.75. After 56 d ad libitum feeding, steers were moved to group pens, ad libitum fed 4 days (ADLIBFED), then fasted 4 days (ADLIBFASTED). This was followed by the 56 d individual restricted feeding, after which, steers were moved to group pens, fed 4 days of restricted DMI (RESTRICTFED), then fasted 4 days (RESTRICTFASTED). There was no difference in lying time (800 and 804 minutes/day, P = 0.85), lying bouts per day (9.8 and 11.4, P = 0.16) and lying duration (99 and 84 minutes/bout, P = 0.18) between low RFI and high RFI cattle, respectively. Fasted animals had less lying time each day (P &amp;lt; 0.05), manifested by fewer lying bouts per day (P &amp;lt; 0.05), that tended (P &amp;gt; 0.05) to be slightly longer than fed animals. Ad lib animals tended (P &amp;gt; 0.05) to have less lying time each day, with a similar number of lying bouts per day, that were slightly (P &amp;gt; 0.05) shorter than restricted animals. Although activity is not affected by RFI level, results suggest that periods of limited feed availability increases activity level.

EARLY TIME RESTRICTED FEEDING IMPROVES HIGH DENSITY LIPOPROTEIN FUNCTION IN GERIATRIC MONKEYS

Ageing conveys the greatest risk for cardiovascular disease (CVD) development, which is the dominant cause of mortality in developed nations. High density lipoprotein (HDL) particles mediate reverse cholesterol transport, are anti-inflammatory and their function predicts CVD. We observed lower plasma cholesterol efflux capacity in geriatric vervet monkeys (p=0.03) when consuming either healthy or Western diets. Adult (n=16) and geriatric (n=19) monkeys were stratified into groups fed Western diet on ad libitum (Ad Lib) or early time restricted feeding (eTFR) schedules. eTRF supplied excess food only between 6am to 2pm. Housing, seasonality and fasting conditions for data and sample collections were equivalent. After 6 months, cholesterol efflux to HDL was greater in eTRF monkeys (p=0.01), with no age by group interaction. Efflux media and plasma was chromatographically separated to confirm labelled cholesterol, and enzymatically measured cholesterol, respectively, was affiliated with HDL particles. eTRF monkeys had higher total plasma cholesterol levels (p=0.03) which was due to greater cholesterol amounts associated with only HDL, and resulted in HDL particles that were larger. eTRF resulted in robustly better HDL function such that measures from geriatric individuals were comparable to younger adults. Additionally, no differences in adiposity was observed in eTRF monkeys. Few interventions are known to raise HDL levels, and more importantly, are confirmed to improve HDL function. Our study is to date the largest, longest, controlled eTRF evaluation in primates and we show that positive biological effects are observable in HDL isolated from both adult and geriatric individuals independently of weight change.

Human Milk and Breastfeeding Outcomes in Infants With Myelomeningocele

In 2015, the study setting instituted an enteral feeding pathway, "PO Ad Lib Feeding to Support Breastfeeding." Many infants admitted to the study setting's newborn/infant intensive care unit with a primary diagnosis of myelomeningocele fall within the setting's enteral feeding pathway's inclusion criteria. The primary objective of this study is to describe the enteral feeding exposure and trends, by type and method, among infants with myelomeningocele. Retrospective descriptive cohort design. Participants were infants with a primary diagnosis of myelomeningocele between 2013 and 2016. The electronic health record was used to collect descriptive data (demographics and daily enteral feeding exposure). Data were analyzed using descriptive statistics. More than 80% (n = 148, 81.3%) of the participants' mothers had a personalized prenatal nutrition consultation and the majority of those mothers (n = 102/118, 86.4%) had a goal to breastfeed for more than 6 months. The majority (n = 144/182, 79.1%) of the study cohort was exposed to mothers' own human milk as their first feed. It is also notable that 80.8% (n = 147) fed at least once directly at the breast; however, infants were also fed by bottle or by feeding tube. It demonstrates that with appropriate evidence-based breastfeeding interventions, mothers having infants with myelomeningocele can expect to feed their infants human milk as well as direct breastfeed. Future research should focus on human milk/breastfeeding outcomes of infants with myelomeningocele post-hospital discharge. Long-term breastfeeding may have an important role in the development of infants with myelomeningocele.

Abstract 2339: High fat diet modulates host immunity and impairs the efficacy of combined radiation and immunotherapy

Abstract Introduction: Obesity is associated with a higher incidence of cancer, increased cancer mortality, impaired responses to radiation therapy (RT), and worse toxicity after RT. Paradoxically, recent studies suggest that obese patients may actually have improved outcomes with immunotherapy (IO), such as programmed death (PD)-1 inhibition, highlighting a link between the host’s metabolic state and immune homeostasis. We therefore hypothesized that a high fat diet could be used to alter the host’s immune response to improve the efficacy of combined RT and IO. Materials/Methods: 12-week old female Balb/c mice were injected with 5x104 syngeneic, luciferase tagged, triple-negative, murine breast cancer cells (4T1). Upon development of a palpable tumor, the mice were randomized to either an ad lib (AL) or high fat (HF), defined as 45% calories from fat, diet. Additionally, each cohort was treated with either diet alone (D), diet plus RT (4 Gy in 3 daily fractions), diet plus PD-1 inhibitor, or a combination of all three (Combo). On day 26, the mice were injected with luciferin and tumor growth was measured using the IVIS bioluminescence in vivo imaging system. At time of sacrifice, primary tumors and spleen were harvested for downstream analysis of immune markers via RNA microarray and flow cytometry. Results: The median overall survival (OS) for animals fed either an HF or AL diet was 36 and 39 days, respectively (NS). The difference in OS however became much more apparent between the individual treatment arms: HF+αPD1 vs AL+αPD1 (37d vs 41.5d, p=0.041); HF+RT vs AL+RT (36d vs 45d, p=0.039); HF+Combo vs AL+Combo (39d vs 48d, p=0.037). HF fed mice also developed metastases sooner than AL fed mice across all arms (D: 22d vs 30d; RT: 24d vs 32d; αPD1: 24d vs 34d; Combo: 27d vs 37d, p=0.047). Microarray analysis of primary tumor tissue revealed an at least 1.5 fold increase expression of cytokines associated with inflammation including IFN-y, TNF-α, IL-17F, IL-22 and IL-1β in the HF diet groups while AL fed mice demonstrated increased expression of the anti-inflammatory cytokines IL-10 and TGF-β. Consistent with the increase in inflammatory markers after a HF diet, flow cytometric analysis of splenocytes revealed a significant decrease in the number of CD4+CD25+Foxp3+ regulatory T cells in the HF group compared with AL fed mice (3.3% versus 7.68%, respectively; p = 0.006). Notably, these Tregs also produced less IL-10 compared with those from AL mice (24.4% vs 54.5%, p=0.037). Conclusions: We demonstrate that a HF diet impairs the efficacy of RT, IO, and combined therapy. This is associated with the development of a pro-inflammatory state demonstrated by increased expression of pro-inflammatory cytokines and a decrease in the number and activity of regulatory T cells. Based on these findings, we conclude that diet can directly impact treatment efficacy and should be considered as a variable in future clinical studies. Note: This abstract was not presented at the meeting. Citation Format: Gregor Manukian, Brittany A. Simone, Tiziana DeAngelis, Kevin Ko, Charles Kivolowitz, Sylvester Jusu, Nicole L. Simone. High fat diet modulates host immunity and impairs the efficacy of combined radiation and immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2339.

S206. Effect of Alcohol on Ad Lib Eating: The Role of Food-Related Reward and Inhibitory Control

Heavy alcohol use is associated with obesity. This may be due to calories in the alcohol beverage itself or to effects of alcohol on food consumption. For example, alcohol may increase food consumption by increasing the value of food rewards, or by decreasing inhibition, which limits food intake.

Harnessing Metabolism with Radiation Therapy to Modulate Host Immunity in the Tumor Microenvironment

Host immunity affects response to immunotherapy (IO) and dictates T cell presence in the tumor microenvironment (TME). Analyses of patient IO response have not accounted for host metabolism, which contributes to chronic inflammation, directly affecting host immunity. As the use of combined IO and radiation therapy (RT) increases, targeting host metabolism is key. We have previously shown that caloric restriction (CR) can augment apoptotic effects of RT and set out to determine if CR could better prime the host for IO+RT therapies. Forty 12-week old female Balb/c mice were injected with syngeneic triple negative murine breast cancer (4T1) cells (5 x 104) and upon formation of a palpable tumor, were assigned to one of four treatment groups: ad lib (AL) fed, 8 Gy RT to the primary tumor (RT), 30% reduction in caloric intake (CR) or CR+RT. Primary tumor tissue was isolated and processed for analysis of immune markers. In the clinic, an IRB-approved feasibility trial using diet (25% CR for 10 weeks) concurrently with RT (60 Gy) for early-stage breast cancer patients was implemented. Serum was collected from patients before RT and after completion of RT and diet. Multiplex ELISA quantified cytokines in sera from trial patients and historical controls receiving RT alone. MicroRNA expression was evaluated in sera. Gene ontology analysis was done on microarray data as well as human sera. T-tests were used to compare expression levels and p ≤ 0.05 was significant. Preclinical and human data both implicated inflammation as a top pathway affected by RT and CR. Cytokine and microRNA data from the preclinical model led to further analysis of STAT3/5. RT alone increased STAT3 and STAT5 (2.5-fold; p=0.04 and 2.1-fold; p=0.03) in primary tumors, while CR alone decreased STAT3 and STAT5 (1.5-fold and 2.4-fold). This decrease remained with CR+RT. Co-expression of pSTAT3 and pSTAT5, which suggests Treg phenotype, was decreased in both the CR and CR+RT arms. Given that pSTAT3 targets miR-21, it was measured as a potential biomarker. MiR-21 was increased in the RT arm compared to AL (3.5-fold; p=0.003) and was decreased in both CR (2-fold; p=0.03) and CR+RT arms (2.5-fold; p=0.01). With respect to T cell function, IFNg was decreased in RT alone, but increased in both the CR and CR+RT treatment arms. Conversely, IL-10 expression was increased after RT alone, but decreased with CR and CR+RT. In human sera, analysis revealed increased memory T cell cytokines (IL-7, IL-15) in CR+RT patients as compared to RT alone and decreased Treg cytokines (IL-10, IL-23). We show that RT creates an immunosuppressive TME via upregulation of pSTAT3, pSTAT5 and IL-10, with a decrease in cytotoxic IFN-g, while CR creates a favorable antitumor TME. A similar cytokine signature seen in CR clinical samples corresponded to decreased levels of miR-21 in serum, making it a potential biomarker for host immunity in this setting. Future trials should explore the use of CR to improve efficacy of IO and RT+IO combination.

Using Ad-Related Network Behavior to Distinguish Ad Libraries

Mobile app ads pose a far greater security threat to users than adverts on computer browsers. This is because app developers must embed a Software Development Kit (SDK), called an ad library or ad lib for short, provided by ad networks (i.e., ad companies) into their app program, and then merge and compile it into an Android PacKage (APK) execution file. The ad lib thus becomes a part of the entire app, and shares the whole permissions granted to the app. Unfortunately, this also resulted in many security issues, such as ad libs abusing the permissions to collect and leak private data, ad servers redirecting ad requests to download malicious JavaScript from unknown servers to execute it in the background of the mobile operating system without the user’s consent. The more well-known an embedded ad lib, the safer the app may be, and vice versa. Importantly, while decompiling an APK to inspect its source code may not identify the ad lib(s), executing the app on a simulator can reveal the network behavior of the embedded ad lib(s). Ad libs exhibit different behavior patterns when communicating with ad servers. This study uses a dynamic analysis method to inspect an executing app, and plots the ad lib behavior patterns related to the advertisement into a graph. It is then determined whether or not the ad lib is from a trusted ad network using comparisons of graph similarities.

How Musical is Australia? A Maverick’s Contemporary Sound Portrait of the Fifth Continent

Violinist/composer Jon Rose builds on How musical is man?, ethnomusicologist John Blacking's classic monograph on the Transvaal Venda people, with the question, ‘How musical is Australia?’. To answer his provocation, in 2002 Rose developed Australia Ad Lib, a website that advocates for a wide concept of what constitutes contemporary music praxis. The Ad Lib website coincides with a time when game-changing innovations to technology and society are on the threshold. Subsequent closure of the site prompts questions concerning the institutional theory of art, the politics of music vis-à-vis marginalised voices, and the perpetuation of cultural cringe. Underpinned by a series of multimedia pieces by Rose, the live show Pannikin features a selection of soloists from Australia Ad Lib—a practical outcome of the website and his research (including lost histories of Australian indigenous and colonial history that he reimagines and resurrects).

Abstract 71: Caloric restriction promotes antitumor immunity through polarization of intratumoral macrophages to M1 phenotype via downregulation of miR-21

Abstract Tumor-associated macrophages (TAMs) are the most abundant inflammatory cells in the breast cancer (BC) tumor microenvironment (TME) and suppress antitumor immune function, contributing to treatment resistance. These M2-polarized macrophages are characterized by fatty acid oxidation as opposed to the glycolytic pathways associated with M1-polarization. MiR-21 is known to affect several metabolic pathways relevant to macrophage polarization. We have previously shown that caloric restriction (CR) can decrease miR-21 expression and increase response to radiation therapy (RT). We propose CR as a means of metabolically reprogramming TAMs toward M1 phenotype to increase efficacy of radiation therapy (RT). To assess the effects of CR in vivo on the composition of the TME in breast cancer, 40 female Balb/c mice were orthotopically injected at 12 weeks old with 4T1 cells and assigned to one of four cohorts: ad lib (AL) fed, 8 Gy RT to the primary tumor (RT), 30% reduction in caloric intake (CR) or CR+RT. MiR-21 levels were significantly decreased in both the CR and CR+RT arms; however, miR-21 was increased in the RT arm. Increased expression of M2 markers was noted in tumors in the RT group as compared to the CR and CR+RT groups, which favored M1 markers. On microarray analysis, the top pathways affected by CR discovered by gene ontology pathway analysis were: inflammation mediated by chemokine and cytokine signaling (31/509 hits), Wnt signaling (21/509 hits) and apoptotic signaling (18/509 hits). Multiple genes related to macrophage polarization were altered by CR including Ccl2 and IL12A. IL-12 is a known target of miR-21, and CR decreases miR-21 expression, therefore we sought to determine if miR-21 may be the mechanism behind the shift in chemokine and cytokine expression in the TME. Cohorts of MMTV-PyMT mice with spontaneously developed breast tumors were bred with a miR-21-/- mouse to generate breast tumors that lack expression of miR-21. These tumors exhibited high levels of IL-12 as well as IL-23 and Cxcl10, all of which are associated with M1 phenotype. MiR-21+/+ tumors exhibited high levels of IL-4 and TGF-β, both associated with M2 phenotype. In addition, peripheral monocytes (PBMCs) were co-cultured with sera from each of the following mice: miR-21-/-, miR-21+/+, AL, RT, CR and CR+RT. M1-polarization was induced by sera from miR-21-/- mice as well as mice receiving CR alone and CR+RT. Ionizing radiation seemed to induce M2-polarization in this manner, as did the miR-21+/+ serum. Taken together, these data suggest that CR can counteract the M2-polarization caused by RT in the TME. This is likely due, in part, to miR-21 downregulation leading to enhanced M1 response. By increasing M1-polarized macrophages and enhancing antitumor immunity, we believe CR is a viable adjunct to standard cytotoxic therapies that could help overcome resistance to chemotherapy and immunotherapy. Citation Format: Brittany A. Simone, Tiziana DeAngelis, Ajay Palagani, Kevin Ko, Larry Harshyne, D. Craig Hooper, Adam P. Dicker, Nicole L. Simone. Caloric restriction promotes antitumor immunity through polarization of intratumoral macrophages to M1 phenotype via downregulation of miR-21 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 71.

Acute Caloric Restriction Leads to Loss of Hypoglycemic Counter-Regulation in Mice following Short-Term Refeeding

Background: Hypoglycemia-associated autonomic failure (HAAF) is a maladaptive failure in glucose counter-regulation known to be caused by recurrent exposure to insulin-induced hypoglycemia. Fasting or caloric restriction is the only natural physiological process which induces prolonged moderate-to-severe hypoglycemia similar to the antecedent of HAAF. In this study we tested the hypothesis that exposure to fasting-induced hypoglycemia can cause HAAF-like symptoms in mice. Methods: Two groups of mice were used, an ad libitum (ad lib) fed group (n=6) and a caloric restriction (CR) group (n=6). CR mice were placed on 60% caloric restriction for 6 consecutive days. Ad lib mice were housed in an identical manner but fed ad libitum during this same period. Following 6 days of restriction, CR mice were given ad lib access to food for 16 h. After the 16 h period of refeeding, both CR and ad lib mice began a 6 h fast which was immediately followed by a hypoglycemic insulin tolerance test (ITT). ITTs consisted of a variable dose of insulin (1.4-1.75 U/kg, IP) which produced similar levels of hypoglycemia in each group (∼45 mg/dL). Results: As expected, mice exposed to 6 days of 60% CR lost ≈30% of their initial body weight, and experienced a gradual fall in blood glucose to an average minimum of 67.9 mg/dL. CR mice displayed a significantly reduced fasting glucose level relative to ad lib mice (102.7 and 171.0 mg/dL respectively, p = 0.0002) as well as reduced serum glucagon levels in response to the ITT (6.0 and 30.4 pg/mL respectively, p = 0.0182). Conclusions: Our results demonstrate that exposure to fasting-induced hypoglycemia produces HAAF-like symptoms in mice following refeeding. These results suggest that HAAF may be a result of an adaptive response to fasting which is inappropriately elicited during exposure to insulin-induced hypoglycemia. Further characterization of the effects of CR on the induction of deficits in glucose counterregulation warrants further investigation. Disclosure D. McDougal: None.

β-cell-specific overexpression of adiponectin receptor 1 does not improve diabetes mellitus in Akita mice.

Adiponectin, a metabolically-active cytokine secreted from adipose tissue, is reported to have anti-apoptotic effects on β-cells as well as anti-hyperglycemic effects through adiponectin receptor signaling. However, the anti-apoptotic effects of adiponectin on β-cells have not been confirmed in established diabetic models, and the anti-hyperglycemic effects and their associated signal cascades remain controversial. To investigate the effects of adiponectin on β-cell protection and its down-stream signaling events, we have generated β-cell-specific rat insulin promoter (RIP)-AdipoR1 transgenic mice (AdipoR1 mice), in which the adiponectin receptor, AdipoR1, is overexpressed in β-cells in a manner synchronous with insulin demand. AdipoR1 mice were then mated with Akita mice, a diabetes model in which β-cell apoptosis results from endoplasmic reticulum (ER) stress. AdipoR1 protein expression and localization in islets from AdipoR1 mice as well as in an AdipoR1-transfected mouse insulinoma cell line were confirmed, as was the activation of both AMPK and Akt in AdipoR1 mice by adiponectin. Nevertheless, there were no significant differences in Ad lib feed and fasting blood glucose levels, or in glucose tolerance tests, between Akita mice [Ins2Akita (C96Y) +/- mouse model] and AdipoR1/Akita and from 4 weeks to 10 weeks of age. Similarly, pancreatic insulin contents of AdipoR1/Akita mice were not significantly different from those in Akita mice from 15 to 20 weeks of age, but they were significantly lower than in wild-type mice. Immunostaining for insulin and subsequent electron microscopy showed that β-cell destruction in AdipoR1/Akita mice was not markedly improved in comparison with that in Akita mice. Serum adiponectin concentrations were confirmed to be extremely high (> 30 μg / ml) compared with the Kd value (0.06 μg / ml) in all mouse groups at 15 to 20 weeks of age. Therefore, although the physiological levels of adiponectin are sufficient to activate AMPK and Akt when AdipoR1 is overexpressed in β-cells, yet adiponectin cannot protect β-cells in Akita mice from ER stress-induced destruction.

A Novel Method to Communicate Po Ad Lib Feeding Goals for the Preterm Infant in the Electronic Medical Record

A Novel Method to Communicate Po Ad Lib Feeding Goals for the Preterm Infant in the Electronic Medical Record

Proses Produksi Iklan Radio di LPP Radio Republik Indonesia Palu

This study aimed to determine types of advertising which is produced in LPP RRI Palu and its production process as well as determine supporting and inhibiting factors of radio advertising production process. The methodology used Action Research based on model characteristics, organizational structure and development goals, system design or scientific development and form of research involvement. The results showed that advertisement was produced in LPP RRI Palu was ad lib, ad spot, and sponsor program. Ad lib production process that was initial briefing, editing script, and broadcast. Ad spot was the initial briefing, creative strategy formulation, drafting, implementation, production of finished advertisement, and evaluation. While ad sponsors program was collaboration of ad spots and ad lib that aired alternately during one program of the event took place. Percentage for ad lib 50% and spot ads 50% per time of ad broadcast. Whereas the program sponsor's only has one or two even in one year there was no broadcast sponsor program. Overall ad impressions per day account for 15% of all broadcast programs.