bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of Ambient Temperature in the Recovery from Major Surgical Stress in Rats
Authors: Powell Romie D , Hetrick Samantha , Nelson Lilly , Schmidt Alex , Brown Justin W.
Published: November 2023

Laboratory animals are routinely surgically instrumented for experimentation and housed at ambient temperatures ranging from 20°C – 26°C during recovery per recommended guidelines. However, rats typically prefer ambient temperatures (Tamb) of ~27 °C (Brown and Le 2011). Rats are often housed at normal room temperature (~22 °C). While this is comfortable to those maintaining them, it may lead to a cold thermal stress for the rats. It is hypothesized that housing rats at ambient temperatures away from their preferred Tamb could lead to a thermal stress which adversely affects surgical recovery. To address this, adult rats (220g–320g) were surgically instrumented with a radiotelemetry probe (DSI), which allows non‐invasive measurement of core temperature (Tc). Also, a cannula (21ga) was inserted into the brainstem to allow microinjection of drugs as part of a separate project. For ≥1 week of recovery, the rats were housed at either 21°C, 24°C, 27°C (control), 30°C, or 33°C Tamb while Tc, food and water intake, and body weight changes were measured. Rats housed at 21°C and 24°C struggled to recover a normal Tc initially. This delayed the onset of normal circadian cycling in the 21°C group. Rats housed at 33°C recovered quickly and then maintained a slightly elevated Tc compared to control (27°C). The 30°C and 33°C groups would reestablish Tc circadian rhythms faster than the control, but the 33°C would struggle to maintain that rhythm near the end of the recovery week. The return of body weight to pre‐surgical levels was delayed in rats housed at the warm Tamb (30°C & 33°C) or the very low Tamb (21°C). The effect of Tamb on daily food and water intake after surgery was not remarkable except for on the 5th day of recovery in the 30°C and 33°C groups when water intake increased. The 21°C and 24°C increased food intake after the 4th day in comparison to the control (27°C). These data suggest that rats maintained at 27°C recovered from surgical stress more readily. These rats returned to pre‐surgical body weight quicker and demonstrated a normal thermoregulatory circadian rhythm earlier than the cold (21 or 24°C) rats. Rats housed at ≥ 30°C may have been exposed to a heat stress which affected weight gain and surgical recovery initially but impaired normal circadian cycling later in the week. Rats housed at 21°C may have been cold stressed which also affected weight gain and thermoregulatory recovery. It is suggested that rats be maintained at their preferred Tamb of 27 °C during the week following surgery to minimize thermal stress and thereby facilitate recovery. This reduction in stress facilitates the return to a normal physiologic state and consequently enables more reliable data collection from these animals.
Sympathetic Neural Control in Chronic Insomnia
Authors: Carter Jason R. , Grimaldi Daniela , Fonkoue Ida T. , Medalie Lisa , Mokhlesi Babak , Van Cauter Eve
Published: November 2023

Heightened sympathetic neural activity and impaired baroreflex function are known contributors to hypertension and cardiovascular risk. Recent epidemiological studies report an association between chronic insomnia and hypertension. In the present study, we examined sympathetic neural and cardiovascular regulation in clinically diagnosed insomniacs and controls. Consistent with the hyperarousal theory of insomnia, we hypothesized that insomniacs would demonstrate higher sympathetic neural outflow and blunted baroreflex control when compared to matched controls. Thirteen insomniacs (40 ± 4 years, 24 ± 1 kg/m2) and 15 matched controls (35 ± 3 years, 26 ± 1 kg/m2; p>0.05) participated in an overnight laboratory polysomnography to exclude obstructive sleep apnea and other sleep disorders, two weeks of at‐home actigraphy, and an overnight laboratory visit with an autonomic function test the subsequent morning. The autonomic function test included simultaneous recordings of heart rate (electrocardiogram), beat‐to‐beat blood pressure (finger plethysmography), and muscle sympathetic nerve activity (MSNA; microneurography) during 10 min supine baseline. Baseline systolic arterial pressure (110 ± 5 vs. 113 ± 4 mmHg; p=0.673), diastolic arterial pressure (69 ± 3 vs. 66 ± 2 mmHg; p=0.424), heart rate (64 ± 3 vs. 59 ± 2 mmHg; p=0.169), and MSNA (23 ± 3 vs. 25 ± 3 bursts/min; p=0.635) were not different between insomniacs and controls. In contrast, spontaneous sympathetic baroreflex sensitivity (BRS) was significantly blunted in insomniacs when compared to controls (−2.1 ± 0.3 vs. −4.3 ± 0.4 bursts/100 heart beats/mmHg; p<0.001). Cardiovagal BRS was not different between groups when assessed as either up‐up (23 ± 4 vs. 25 ± 6 ms/mmHg; p=0.843) or down‐down (25 ±5 vs. 25 ± 5 ms/mmHg; p=0.955) sequencing methods. In conclusion, insomniacs demonstrated similar levels of MSNA, but substantially blunted sympathetic BRS, at rest when compared to matched controls. These findings support growing evidence of increased risk of hypertension with chronic insomnia.

Support or Funding Information

Supported by Merck Investigators Studies Program.
Sleep Restriction Degrades Local Immune Response of Experimental Wounds
Authors: Wilson Marques A , Young Andrew J , Montain Scott J , Smith Tracey J
Published: November 2023

The suction blister model creates experimental skin wounds and permits evaluation of local immune response and wound healing. Sleep restriction negatively affects systemic immune function, e.g., natural killer cell activity, suppressed interleukin‐2 production and increased levels of circulating proinflammatory cytokines; however, the impact of sleep restriction on local immune response at experimental wound sites has not been described. This cross‐sectional study assessed the effect of sleep restriction on wound exudate cytokine concentrations after skin blister formation. Up to 8 blisters (~30 mm2) were induced viasuction on participant's forearm either after 5 nights of adequate sleep (AS,7–9‐h sleep per night confirmed via activity monitors) or on the second day of a 3‐d sleep restriction period (SR, monitored in laboratory with ~2‐h sleep pernight). The top layer of blisters was removed to reveal up to 8 wounds. Wound exudate was sampled at 4, 7 and 24‐h post blistering to assess cytokine response via multiplex assay and area‐under‐the‐curve with respect to baseline cytokine concentrations (AUCi) was calculated. Skin barrier restoration was assessed by transepidermal water loss, measured daily until values were within 90% of unbroken skin. Data, analyzed with mixed models analysis of variance and independent samples t‐tests, is reported for the sixteen (AS: 23 ± 5 years [mean±SD]) and twenty (SR: 21 ± 4 years) study completers. Sleep duration in the 5‐days leading up to either the blister procedure (AS) or the sleep restriction period (SR) was not different between groups (7.99 ± 0.5 hrs·d−1 and 7.71 ± 0.67 hrs·d−1, respectively, P = 0.18). Wound exudate concentrations of IL6, IL8, MIP1α, MIP1βand TNFα significantly increased over time for both AS and SR (p < 0.05);and, concentrations of IL6 and IL8 were higher for AS compared to SR at 7‐h (P= 0.03 and 0.004, respectively). The AUCi was higher for AS compared to SR with regard to IL8 (log10, 5.1 ± 0.2 and 4.9 ± 0.2, respectively, P = 0.03), but not other cytokines. Skin barrier restoration was shorter for AS compared to SR: 4.2 ± 0.9 and 5.0 ± 0.9 days, respectively (P = 0.02). These data support the hypothesis that relatively modest sleep disruption degrades immune response at the wound site and delays wound healing. Interventions to ameliorate decrements in immunefunction and wound healing in response to sleep restriction deserve exploration.

Disclaimer: The views expressed in this abstract are those of the authors and do not reflect the official policy of the Department of Army, Department of Defense, or the U.S. Government.

Support or Funding Information

Funded by the Medical Research and Materiel Command, Aberdeen Proving Ground, Maryland.
Developmental Exposure to Indoor Flame Retardants Produces Diabetic Phenotype in Adult Female C57BL6 Mice
Authors: Kozlova Elena , Krum Julia Meagan , Basappa Karthik , Uddin Syed , Curras‐Collazo Margarita C
Published: November 2023

Polybrominated diphenyl ethers are a group of flame‐retardant organohalogen pollutants that act as endocrine and neuroendocrine disrupting chemicals. In humans, exposure to brominated flame retardants or other environmentally persistent organic pollutants (POPs) such as polychlorinated biphenyls (PCBs) has been associated with increasing trends of diabetes and metabolic disease (Gray et al. 2013; Baker et al. 2013; Lim et al, 2008; Lee et al, 2006). POPs may disrupt metabolic homeostasis through chronic low‐level exposure and bio‐accumulation in liver, adipose tissue and brain (Zhang et al, 2016; Turyk et al, 2015). However, the effects of PBDEs on metabolic processes are poorly understood. In this study we examined the effects of gestational and lactational exposure to the industrial PBDE mixture DE‐71 on glucose tolerance test (GTT), insulin tolerance test (ITT), and liver lipid content in C57BL/6 female offspring in adulthood (128–147 days; 17.1–22 g). We also examined intestinal permeability since intestinal‐derived bacterial products entering the bloodstream may cause inflammation, liver disease that may contribute to obesity and diabetes (Turner, 2009). Mice were exposed to environmentally relevant doses of PBDEs for 10 weeks (pre‐conception: 4 weeks; gestation: 3 weeks; lactation: 3 weeks). Daily dosing consisted of low dose (0.1 mg/kg/d), high dose (0.4 mg/kg/d), or corn oil vehicle (control) via ingestion of infused corn flakes. GTT results revealed that blood glucose levels normalized to fasting levels were significantly elevated at 30 and 60 min post injection of glucose (2.0 g/kg) in LD mice relative to oil vehicle controls (p<0.0001, n=20). ITT results showed a significantly decreased sensitivity to insulin (0.5 U/kg) at post‐injection time points from 15–120 min for LD and 15–90 min for HD female offspring (p<0.05 to 0.0001, n=19). Mean levels of liver lipids were significantly elevated in LD female offspring compared to controls (p<0.05, n= 18). To determine if PBDE‐exposed offspring showed altered intestinal permeability, mice were orally gavaged with 60 mg/kg FITC‐dextran (mol. wt. 3,000–5,000; FD‐4, Sigma) and fluorescence measured in the blood 4 hr later by fluorimetry (excitation 490 nm, emission 530 nm). PBDE at HD had an apparent effect on elevated plasma FITC levels but the effects were not significant indicating that PBDEs may not produce their effects via changes in intestinal barrier integrity. These results indicate that mice perinatally exposed to PBDEs (0.1 mg/kg dose) display abnormal glucose and lipid metabolic parameters indicative of a diabetic phenotype.

Support or Funding Information

Supported by University of California COR grant and UCMEXUS.
Contribution of POMC and AgRP neurons to the control of metabolic autonomic nerve activity by leptin
Authors: Bell Balyssa , Morgan Donald , Rahmouni Kamal
Published: November 2023

Central action of the adipocyte‐derived hormone leptin is critical for metabolic regulation. Leptin promotes energy expenditure by increasing sympathetic nerve activity (SNA) to thermogenic brown adipose tissue (BAT). Leptin also increases hepatic SNA and parasympathetic nerve activity (PNA) to modulate glucose production by the liver. We previously demonstrated the importance of the hypothalamic arcuate nucleus (ARC) in underlying leptin‐induced increases in BAT SNA as well as liver SNA and PNA, but the contributions of specific neuronal populations within the ARC are not fully understood. Here, we tested the hypothesis that proopiomelanocortin (POMC) and agouti‐related peptide (AgRP) neurons of the ARC mediate regional SNA and PNA responses evoked by leptin. To test this, we used direct multifiber nerve recording to assess the effects of intracerebroventricular (ICV) leptin (2 μg) on regional nerve activity in mice lacking leptin receptors on POMC (POMCCre/LepRfl/fl) or AgRP neurons (AgRPCre/LepRfl/fl). We observed a partially blunted BAT SNA response to ICV leptin in both POMCCre/LepRfl/fl (148±29%, p<0.05) and AgRPCre/LepRfl/fl mice (172±62%, p<0.05) as compared to littermate controls (327±62%), suggesting a distributed control of BAT thermogenesis by leptin across neuronal populations. Conversely, the hepatic SNA response to leptin was significantly reduced in AgRPCre/LepRfl/fl mice (76±21%, p<0.05), but not in POMCCre/LepRfl/fl mice (120±49%) relative to controls (196±36%). However, the hepatic parasympathetic response to leptin was completely abolished in mice lacking leptin receptors on either POMC (−17±17%) or AgRP (20±15%) neurons vs control mice (108±24%, p<0.05), indicating that both of these populations are required for leptin to suppress hepatic glucose production. Together, these findings support a critical role for hypothalamic neurocircuitry in dissociating the regional autonomic effects of leptin, in particular with respect to thermogenic control and glucose homeostasis.
SPAK Kinase Modulates Energy Expenditure and Thermogenesis in Mice Fed a High‐fat Diet
Authors: Torre‐Villalvazo Ivan , Cervantes‐Perez Luz Graciela , Noriega Lilia Guadalupe , Jimenez José Victor , Uribe Norma , Torres Nimbe , Bobadilla Norma Araceli , Tovar Armando , Gamba Gerardo
Published: November 2023

Body weight regulation and glucose homeostasis are controlled by central and peripheral mechanisms. In the central nervous system, hypothalamic nuclei trough neuronal axes that release GABA modulate energy expenditure and thermogenesis and the peripheral glucose metabolism is controlled in part by pancreatic insulin secretion from β‐cells. Interestingly, the intracellular chloride concentration ([Cl−]i) is involved in the modulation of both processes, the GABA signaling and insulin secretion. The activity of the electroneutral cation coupled chloride cotransporters (CCC) is critical to define the [Cl−]i. Phosphorylation of the CCC members promotes Cl− influx, while dephosphorylation promotes Cl− efflux. The kinase that phosphorylates the CCCs is known as SPAK (STE20‐proline alanine rich kinase). The SPAK activity modulates the [Cl−]i by controlling the activity of the CCCs. In this regards, the homozygous SPAK‐Knockin mice lacking the threonine 243 of the T‐loop required for its activation (SPAKT243A/T243A) exhibit no SPAK activity and thus a marked decrease in CCCs phosphorylation. Here, to analyze the role of SPAK in metabolic homeostasis we fed wild type and SPAKT243A/T243A mice a high‐fat diet (HFD) for 17 weeks to evaluate weight gain, metabolic variables, energy expenditure, glucose tolerance, as well as, liver, pancreatic and adipose tissue histology and histochemistry. Food consumption was similar among groups during the feeding period. The wild type mice, however, gained 15.1 ± 0.8 g, whereas the SPAKT243A/T243A mice gained only 10.2 ± 2.0 g (p<0.01). At the end of the study, in contrast to wild type mice, SPAKT243A/T243A mice exhibited significantly lower fat mass, cholesterol, triglycerides and leptin levels, as well as improved insulin secretion and glucose tolerance. SPAKT243A/T243A mice did not develop hepatic steatosis and pancreatic islets hypertrophy as wild type did. The enhanced metabolic function of SPAKT243A/T243A mice was associated with increased whole body oxygen consumption and augmented fat oxidation. The molecular mechanisms responsible for this phenotype involve improved pancreatic β‐cell function, augmented skeletal muscle mitochondrial activity and increased brown adipose tissue UCP1‐mediated thermogenesis. Our data show that the absence of SPAK activity in mice is associated with an obesity resistant phenotype when exposed to HFD. These observations suggest that modulation of SPAK activity could be a potential new therapeutic avenue for obesity and glucose metabolism disorders.

Support or Funding Information

Grant No. 23, of the Fronteras de la Ciencia program of Mexican Council for Science and Technology (CONACYT)
Role of H3K36 methylation in cell cycle control and nutrient stress response
Authors: Strahl Brian
Published: November 2023

H3K36 methylation (H3K36me) is a co‐transcriptional modification critical for the maintenance of chromatin structure and the fidelity of transcription. However, the extent to which the control of transcriptional fidelity by Set2/H3K36me broadly impacts cellular functions such as cell cycle control and stress response is not known. In my presentation, I will describe our recent work into the role of the H3K36 methyltransferase Set2. Our studies show that Set2 is not only cell cycle regulated, but also required for proper cell cycle progression and nutrient stress response – findings that appear to be conserved in humans. We propose that the ability of Set2/H3K36me to reinforce chromatin structure and prevent pervasive anti‐sense/cryptic transcription is required for the proper regulation of highly timed transcription programs such as those found in the cell cycle and nutrient signaling.
Resolution of head‐on replication‐transcription conflicts in bacteria
Authors: Merrikh Houra , Hall Ashley , Lang Kevin
Published: November 2023

The DNA replication and transcription machineries function at the same time and on the same template, inevitably encountering each other. These encounters generate conflicts that if not resolved, can have dire consequences for the cell including replisome stalling, increased mutagenesis, and DNA breaks. Conflicts can occur in either the co‐directional (leading strand genes) or head‐on (lagging strand genes) orientation. Although both types of encounters can stall the replisome and lead to breaks, head‐on conflicts have much more detrimental consequences for both replication and genome stability. In the Merrikh lab, in addition to investigating the consequences of conflicts on replisome structure, mutagenesis, and evolution, we are focused on identifying essential conflict resolution mechanisms. We use bacterial model organisms for our research because they are easy to genetically manipulate, but also have a single chromosome with a single origin of replication, allowing us to easily isolate the effects of gene orientation. We have recently identified several new conflict resolution factors, without which, a single severe head‐on conflict can lead to significant survival defects, replication stalling, and problems with gene expression. In this talk, I will discuss our recent progress on the topic and share new data from our lab regarding these conflict resolution factors in bacteria. Overall, our findings indicate that for cells to survive expression of genes in the head‐on orientation, they have to possess specialized conflict resolution factors. The evolutionary conservation of these factors, together with the presence of lagging strand genes, suggests that head‐on replication‐transcription conflicts routinely create a problem that must be resolved by cells across all domains of life.
The Evf2 enhancer lncRNA links chromosomal interactions and interneuron diversity
Authors: Kohtz Jhumku , Cajigas Ivelisse , Swyter Kelsey R , Bastidas Monique , Chakraborty Abhijit , Morris Elizabeth , Ay Ferhat
Published: November 2023

Understanding how genes are selectively regulated during development is a major goal in biology. While DNA enhancers selectively regulate genes at a distance, mechanisms regulating long distance enhancer interactions are only beginning to be understood. Our previous work reported transcriptional activity by Evf2, an enhancer long non‐coding RNA (lncRNA) transcribed from the Dlx5/6 ultraconserved enhancer (Dlx5/6ei)1. Evf2 regulates adjacent chr6 genes Dlx5/6, recruiting both activators and repressors to Dlx5/6ei, and forms a large ribonucleoprotein complex (Evf2‐RNP, ~79 proteins) that contains the DLX1 homeodomain transcription factor and chromatin remodelers2,3,4. Promiscuous interactions between Evf2 and chromatin remodelers reveal RNA‐mediated direct inhibition of BRG1(SMARCA4) ATPase and remodeling activities4. Thus, enhancer RNAs can regulate enhancer activity by providing a scaffold for multiple promiscuous RNA binding proteins, and directly regulating enzymatic activity. Here, we show evidence supporting that Evf2 controls chromosome‐wide enhancer interactions and histone modifications, selectively regulating genes located in a ~27Mb region of chr6 (Evf2‐chr6 targets). These data support the idea and that while Evf2 plays a major role in chromosome topology, only a subset of Evf2‐dependent enhancer interactions contribute to gene regulation. Evf2 loss, truncation, and rescue distinguish between the roles of the Evf2‐5′ ultraconserved enhancer‐containing region and trans/cis mechanisms that regulate Evf2‐chr6 targets. Together, this work identifies a novel network of interneuron subtype gene regulation that links enhancer lncRNA directed chromosomal interactions and interneuron diversity in the embryonic mouse brain.

Support or Funding Information

NIMH R01MH094653 to J.D.K
Metabolic Disease Symptoms are Reduced in C57BL/6J Mice Fed High‐fat Diets Supplemented with Oak Tannins
Authors: Shay Neil F , Luo Ting F , Sturm Marlena F
Published: November 2023

Previous work has demonstrated that moderate red wine consumption, and intake of some bioactive compounds in wine including quercetin and resveratrol correlate with a reduced incidence of metabolic disease symptoms. The aging of wine in oak barrels adds to the polyphenolic content of wine by introducing oak tannins to the wine. Tannins have been suggested to be related to the hypothesized beneficial effect of wine against the progression of cardiovascular and metabolic disease.

The objective of the present study is to investigate the influence of oak tannin intake on the symptoms of metabolic syndrome induced by the consumption of a high fat diet fed C57BL/6J (C57) mice. C57 mice were fed low fat (LF), high fat (HF), and HF containing 0.16% (w/w) of oak tannin powder (HF+OT) for 10 weeks. HF + OT intake significantly reduced body weight (42.4g±1.2g [HF] vs. 38.2g±1.4g [HF+OT]). Glucose homeostasis was regulated by OT intake, e.g., serum resistin levels were lower in HF+OT‐fed mice, compared to HF‐fed mice (374±11 pg/mL in HF vs.301±10 pg/mL). Further, hepatic lipid accumulation was lower in the HF+OT‐fed group (p<0.05). To further understand the underlying mechanisms of how OT intake regulates glucose and lipid metabolism, hepatic gene expression related to lipid, glucose, and xenobiotic metabolism was evaluated. Relative expression of Gstm2 was significantly decreased in the HF+OT group (p<0.05). Scd1 gene expression was increased in the HF‐fed group, and reduced to a value resembling the LF‐fed group in HF+OT fed mice. Nqo1 and Cyp2b10 gene expression was decreased with OT intake. Serum levels of the pro‐inflammatory cytokine MCP‐1, was marginally reduced HF+OT‐ vs. HF‐fed mice (p=0.0877).

A follow‐up study was completed to determine whether the consumption of oaked vs. unoaked red wine can influence metabolic syndrome parameters in HF‐fed mice. In a ten‐week trial, mice were fed LF or HF diets, and HF diets with an oaked wine concentrate (HF+OWC) or unoaked wine concentrate (HF+UWC). Intake of both the HF+OWC and HF+UWC reduced body weight gain, compared to HF‐fed mice. Hepatic gene expression evaluation of Lipe, Cyp2b10, and Gsta showed more significant regulation of these mRNAs with intake of OWC vs. UWC.

We conclude that intake of oak tannins have the potential to act as an agent to ameliorate symptoms of metabolic syndrome.

Support or Funding Information

Desert Heart Foundation
Muscadine Grape Extract Inhibits Proliferation and Migration of Triple Negative Breast Cancer Cells in Association with a Reduction in TWIST
Authors: Collard Marianne , Gallagher Patricia E. , Tallant Ann
Published: November 2023

Muscadine grape (V. rotundifolia) seeds and skins are used as nutraceuticals for their anti‐oxidant, anti‐inflammatory and anti‐cancer properties. However, the effect of treatment with muscadine grapes on different types of cancers, including breast cancer, is limited. Triple negative breast cancer is a subtype of breast cancer which is characterized by the lack of estrogen receptors, progesterone receptors and over‐expression of the epidermal growth factor receptor, limiting targeted treatment for women with the disease. In addition, triple negative breast cancer which disproportionally affects ethnic minorities and younger women has a high propensity to metastasize, rendering it one of the more aggressive breast cancer subtypes. The goal of this project was to determine whether a proprietary muscadine grape extract (MGE; from Piedmont Research & Development Corp.) reduces the growth and metastatic progression of triple negative breast cancer. A dose‐ and time‐dependent decrease in proliferation was observed following MGE treatment of murine 4T1 cells (83.3% inhibition at 25 μg/mL for 24 h; n=3, p < 0.01) and human MDA‐MB‐231 (49.4% inhibition at 25 μg/mL for 48 h; n=3, p < 0.001), BT‐549 (46.0% inhibition at 25 μg/mL for 48 h; n=3, p < 0.001) and BT‐20 (43.6% inhibition at 25 μg/mL for 48 h; n=3, p < 0.001) triple negative breast cancer cells as compared to untreated cells. MGE also reduced the migration of triple negative breast cancer cells measured using a scratch wound assay. The significant reduction in migration of 4T1 (220.9 ± 16.6 μm vs 44.0 ± 6.2 μm at 25 μg/mL for 12 h; n=3, p < 0.001), MDA‐MB‐231 (549.2 ± 53.5 μm vs 268.3 ± 41.5 μm at 25 μg/mL for 18 h; n=3, p < 0.01) and BT‐549 (424.4 ± 40.7 μm vs 140.3 ± 33.6 μm at 25 μg/mL for 36 h; n=3, p < 0.01) cells by MGE was dose‐ and time‐dependent as well as independent of proliferation.

The transcription factor TWIST, which is involved in the epithelial‐to‐mesenchymal transition, is critical in the development of metastasis due to the increased migration and invasion of cancer cells exhibiting the mesenchymal phenotype. MGE significantly reduced TWIST expression at a concentration of 20 μg/mL in 4T1 triple negative breast cancer cells with a maximal decrease of 64.6% compared to untreated cells after 12 h of treatment (n = 5; p < 0.05). Furthermore, phospho‐ERK1/2, a MAP kinase which positively regulates TWIST, was reduced approximately 2‐fold (n = 5; p < 0.05) after 10 h of 20 μg/mL MGE treatment or greater than 3‐fold (n = 6; p < 0.05) after 12 h. Taken together, these results suggest that MGE reduces ERK1/2 activities to down‐regulate TWIST and inhibit the migration of triple negative breast cancer cells, and that treatment with MGE may be an effective therapeutic to decrease the progression of triple negative breast cancer and prevent metastasis at distant sites.
Monomeric cocoa procyanidins enhances β‐cell function by increasing mitochondrial respiration
Authors: Tessem Jeffery Sivert , Rowley Thomas J. , Bitner Benjamin F. , Ray Jason D. , Smithson Andrew T. , Hansen Jason M , Bikman Benjamin T. , Neilson Andrew P
Published: November 2023

A hallmark of type 2 diabetes (T2D) is β‐cell dysfunction and the eventual loss of functional β‐cell mass. Therefore, mechanisms that can improve β‐cell function could be used as a treatment for the millions of individuals with T2D. Recent studies have shown that monomeric, oligomeric and polymeric cocoa derived procyanidins have vastly different effects on obesity, insulin resistance and glucose tolerance during high fat feeding. Based on these previously published data, we hypothesized that cocoa derived procyanidins may have beneficial effects on improving β‐cell function. 832/13 INS‐1 derived β‐cells and primary rat islets cultured with monomeric cocoa procyanidins demonstrated enhanced glucose stimulated insulin secretion (GSIS), while cells cultured with total cocoa extract, oligomeric procyanidins or polymeric procyanidins demonstrated no improvement. The increased GSIS observed with culture in the presence of monomeric procyanidin fractions corresponded with enhanced mitochondrial respiration, suggesting improvements in β‐cell fuel utilization. We demonstrated upregulation of electron transport chain components Complex III, IV and V after culture with monomeric procyanidins. Monomeric procyanidins also resulted in an increased cellular redox state and increased glutathione concentration, which corresponds with Nrf2 nuclear localization. We propose a model by which cocoa derived monomeric procyanidins enhance the cellular redox state, allowing Nrf2 to migrate to the nucleus and enhance expression of key mitochondrial genes that results in enhanced GSIS and improved β‐cell function. These results suggest that cocoa derived monomeric procyanidins could be leveraged as a treatment modality for patients with T2D.

Support or Funding Information

BYU MEG Grant to JST, Diabetes Action Research and Education Grant to JST, BYU ORCA Grant to JDR and BFB
Altered Postprandial Lipemia and Fatty Acid Handling in Obese vs. Normal‐Weight Men Is Rectified by Reducing Dietary Fat Load: A Dose‐Response Trial
Authors: Vors Cécile , Gabert Laure , Drai Jocelyne , Louche‐Pélissier Corinne , Laville Martine , Vidal Hubert , Michalski Marie‐Caroline
Published: November 2023

Background

Postprandial hyperlipemia and altered dietary lipid beta‐oxidation are now recognized as metabolic risk factors in obesity that are directly associated with dietary fat intake. The amount and size of chylomicrons are known to impact their clearance and thus possibly modulate the final partitioning of dietary fatty acids (FA) between β‐oxidation and storage, which is altered in obesity. However, the detailed impact of ingested fat amount on postprandial lipemia and dietary FA fate in obese subjects remains to be elucidated.

Methods

In a randomized crossover study, eighteen healthy normal‐weight (NW) and obese men ingested meals containing 10g or 40g of fat (typical breakfast fat contents) labeled with a mix of 13C‐triglyceride tracers. Chylomicron triglyceride content and size were measured during 8h of digestion. Plasma concentrations of 13C‐palmitate and 13C‐oleate were also measured during 8h in parallel with their fecal loss (72h‐stool collection). 13CO2 breath‐test coupled to indirect calorimetry measures during 8h allowed to calculate exogenous lipid fate.

Results

Chylomicron triglycerides increased in all subjects according to ingested fat amount (P<0.01). After 40g of fat, obese men had delayed postprandial lipemia compared to NW men (PtimexBMI<0.0001) especially at 5–8h post‐breakfast (P<0.01), with different variations in chylomicron size (PtimexBMI<0.01). This was associated with a lower appearance of tracers in plasma in obese men (P<0.01 for AUC 0–5h vs. NW), and a tendency to higher fecal excretion of 13C‐oleate (P=0.1 vs. NW) after 40g of fat. However after 10g of fat, chylomicron TG and size, and tracer kinetics and fecal loss, were similar regardless of BMI. Finally, the impact of fat amount on exogenous lipid β‐oxidation was different according to BMI (PdosexBMI<0.01): 39.6% of ingested lipids were β‐oxidized after 40g of fat in obese (vs. 45.1% in NW) but this was increased to 53.1% in obese for 10g of fat (vs. 49.7% in NW subjects; P<0.001 vs. 40g).

Conclusions

Postprandial lipemia profile after a realistic high‐fat load is altered in obese subjects. Although the reduction of ingested fat amount seems to normalize the postprandial fate of dietary lipids in obese men, further research is needed to understand mechanisms of altered postprandial lipid metabolism in obesity.

Support or Funding Information

The Study was funded by CNIEL (French Dairy Interbranch Sector) and INRA (French National Institute for Agricultural Research).
Intake of up to Three Eggs per Day is Associated with Improvements in Markers of HDL Function and Antioxidant Capacity in a Young, Healthy Population
Authors: DiMarco Diana M , Norris Gregory H , Millar Courtney L , Blesso Christopher N , Fernandez Maria Luz
Published: November 2023

Recent evidence suggests that the composition and function of HDL particles may be more important than HDL cholesterol concentration when assessing risk for cardiovascular disease (CVD). HDL is involved in reverse cholesterol transport, the process by which excess cholesterol is removed from tissues and returned to the liver for biliary excretion. In addition to transporting cholesterol, HDL particles carry antioxidant proteins, enzymes, and carotenoids, all of which help protect against oxidative damage to both HDL and LDL particles. Eggs contain cholesterol and carotenoids and therefore may impact CVD risk by modulating lipoprotein composition and function. The goal of this research was to determine the impact of intake of up to 3 eggs/d on LDL and HDL particle size, function, and associated antioxidants in a young, healthy population. Thirty‐eight healthy individuals (19 men/19 women, 24.1 ± 2.2 y, BMI 24.3 ± 2.5 kg/m2) completed this 14‐week crossover dietary intervention. Qualifying participants underwent a 2‐week washout (0 eggs/d), followed by intake of 1, 2, and 3 eggs/d for 4 weeks each. Following each dietary period, fasting plasma and serum were collected for analysis of lipoprotein particle size and subfractions by nuclear magnetic resonance and concentrations of plasma apolipoproteins and carotenoids (lutein and zeaxanthin). Activities of plasma cholesteryl ester transfer protein (CETP) and lecithin‐cholesterol acyltransferase (LCAT) were assessed using fluorometric kinetic assays. Lastly, serum paraoxonase‐1 (PON1) activity toward phenyl acetate was measured using an in‐house kinetic assay. Compared to 0 eggs/d, intake of 1–3 eggs/d led to increased large LDL and large HDL particle concentrations (p < 0.05), a 9–20% increase in plasma apoA‐I (p < 0.05), and increased LCAT activity (p < 0.05). Furthermore, as compared to 0–1 eggs/d, intake of 2–3 eggs/d resulted in an 11% increase in apoA‐II (p < 0.05) and a 20–31% increase in plasma lutein + zeaxanthin (p < 0.05). Lastly, as compared to lesser quantities of eggs, intake of 3 eggs/d led to increased serum PON1 activity (p < 0.05). In contrast, egg intake had no impact on plasma CETP activity. In summary, intake of 1 egg/d appears sufficient to increase large LDL particle concentration and biomarkers associated with HDL function. However, intake of 2–3 eggs/d supports greater improvements in markers of HDL function and antioxidant capacity. Overall, intake of up to 3 eggs/d improves these biomarkers associated with lipoprotein function and CVD risk in a young, healthy population.

Support or Funding Information

This research was supported by an award to MLF from the Esperance Family Foundation and a grant to DMD from the Egg Nutrition Center.
How to create interactive digital resources that result in real learning outcomes
Authors: Rea Paul , Krebs Claudia
Published: November 2023

Over recent years, technological advances and digital tools in anatomical education have been rapidly developing. Indeed, the way our students learn and access educational resources has changed significantly over the years. This is also reflected in the surge to the market of digital educational material, and not always validated by professional anatomists.

Through an International Anatomy Education Hub, we have combined our skillsets in the field of digital anatomy from anatomy and surgical staff, from four leading universities across the globe. We aim to introduce a variety of tools and software that we as educators, and student users, could use in generating educationally validated, curriculum specific, digital anatomical training materials.

Based on human‐computer interaction research and theories, evidence‐based guides to building effective educational content in the digital environment will be discussed. It will also highlight strategies for involving the students as co‐creators of educational materials, designing assessment tools, and will present feedback from the end user.

Simple and effective ways to build local, curriculum specific content, in an open source format, will be presented, as well as the benefits of international collaborations. This will show how local digital focused activities can benefit the global anatomical education community.
The Influence of Dietary Micronutrients Upon the Risk of ‘Pouchitis’‐like Inflammation in Genetically Susceptible Hosts
Authors: Pierre Joseph F. , Hinterleitner Reinhard , Bouziat Romain , Hubert Nate , Miyoshi Jun , Jabri Bana , Chang Eugene B
Published: November 2023

Ulcerative colitis (UC) is a subset of inflammatory bowel disease (IBD) characterized by inflammation in the colon, but not small bowel. In medically refractory UC, the ultimate treatment is total proctocolectomy and surgical creation of an ileal pouch. Unfortunately, many IBD patients develop inflammation within the pouch, termed ‘pouchitis’. Gut dysbiosis, host genetics, and environmental triggers are implicated as causative factors in IBD, yet mechanisms are poorly understood. Following observations that diet may influence inflammatory risk, we employed our newly established murine model of ‘pouchitis’ – using surgically created ileal self‐filling loops ‐ to investigate the role of dietary micronutrients upon disease risk in genetically susceptible hosts. C57B/6 WT and IL‐10−/− mice were randomized to Control (CON) or Antioxidant (AOX) diet ‐ containing pharmacologically relevant levels of 4 micronutrients – for two weeks following surgical recovery. There were no differences in body weight change or survival; however, IL‐10−/− CON animals had significantly larger lymphoid organs compared with IL‐10−/− AOX or WT mice on either diet. Examination of IL‐10−/− CON loop mucosa demonstrated histological inflammation characterized by goblet cell depletion, increased tissue MPO levels, and elevated IFN‐γ, TNFα, and IL‐17α expression, which was attenuated with AOX. There were no difference in luminal IgA between diets in eiher genotype; however, compared with IL‐10−/− CON mice, AOX mice had significantly greater CD45+CD4+FOXp3+ Treg cells and fewer CD45+CD4+RORg T cells, without changes in CD45+CD4+T‐bet cells. 16S analysis demonstrated AOX increased the Bacteriodetes‐to‐Firmicutes ratio and increased alpha diversity compared with CON diet in both genotypes. Importantly, convention alization of germ‐free IL‐10−/− with CON derived microbes, but not AOX microbes, recapitulated the inflammatory phenotype. These experiments demonstrate the ability of common dietary components to mitigate intestinal inflammatory disease in genetically susceptible hosts and highlight the role of microbial populations in disease pathogenesis. Using this model, further elucidation of immune‐microbial compartment set points that occur under specific dietary components may help us understand, and eventually avoid, dietary factors that result in recurrent inflammatory episodes in susceptible pouchitis patients.

Support or Funding Information

This work was supported by NIH NIDDK F32DK105728‐01A1, P30DK42086.
Determination of CYP2A6 Activity in the US Population by Use of a Caffeine Metabolite Ratio in Spot Urine Samples: NHANES 2009–2010
Authors: Pao Ching‐I , Sternberg Maya R. , Rybak Michael
Published: November 2023

Cytochrome P450 enzyme CYP2A6 is a phase I enzyme expressed primarily in the liver, accounting for approximately 4% of all P450 content. CYP2A6 plays a role in the metabolism of caffeine, nicotine, and several drugs used in the treatment of cancer and infectious diseases. CYP2A6 polymorphisms give rise to inter‐individual differences in CYP2A6 activity, and these differences can be further influenced by dietary and lifestyle factors. Lower CYP2A6 activity has been associated with a reduced risk of various cancers, and differences in CYP2A6 activity can influence smoking behavior and likelihood of cessation. Caffeine metabolite ratios (MR) have been developed to assess CYP2A6 activity, and the urine MR of 1,7‐dimethyluric acid (17U) to paraxanthine (17X) (i.e., 17U/17X) has been used to determine CYP2A6 activity. Additionally, the MR of 17U to 1‐methylxanthine (1X) (i.e., 17U/1X) has shown promise in less controlled settings where regular dietary caffeine intake and spot urine samples were used. In this study we described CYP2A6 activity in the US population ≥6 y by selected demographic, lifestyle and physiologic variables. We used spot urine molar concentrations of 17U and 1X from the cross‐sectional NHANES 2009–2010 to calculate CYP2A6 activity by use of the MR 17U/1X. We excluded the individuals with evidence of insufficient caffeine exposure to reliably calculate the CYP2A6 MR and subjects taking prescription medications known to interfere with caffeine metabolism. We performed bivariate comparisons of unadjusted geometric mean (GM) CYP2A6 MR for categorical variables (n=2197), and used Spearman correlation to examine the relation of CYP2A6 MR with continuous variables. CYP2A6 activity was higher in females vs. males (Wald F P <0.0001), highest in Hispanics, followed by non‐Hispanic whites and lowest in non‐Hispanic blacks (Wald F P <0.0001), and increased with age (Spearman ρ = 0.41, P <0.001). CYP2A6 MR was higher in individuals with evidence of inflammation (serum CRP ≥5 mg/L vs. <5 mg/L; Wald F P <0.0001), diabetes (yes vs. no based on a physician's diagnosis; Wald F P = 0.0025), and impaired kidney function (CKD 3–5 vs. CKD 1–2 and normal; Wald F P <0.0001). CYP2A6 MR was lower in smokers vs. non‐smokers (serum cotinine ≤10 ng/mL vs. >10 ng/mL; Wald F P <0.0001). CYP2A6 activity was positively associated with BMI (Spearman ρ = 0.36, P <0.0001) and was negatively associated with leisure physical activity (Spearman ρ = −0.15, P = 0.0003). The associations that we observed for CYP2A6 activity with age, sex, and race‐ethnicity, smoking status, inflammation, and physical activity were generally consistent with observations from other studies. We believe that our study is the first to report estimates of CYP2A6 activity obtained from spot urine MRs in a representative sample of the US population.
Combination of Quercetin and Green Tea Extract Increased Plasma Epicatechingallate and Decreased Urine Epigallocatechin and Epicatechin Concentrations
Authors: Henning Susanne M , Aronson William J , Wang Piwen , Wang Jieping , Lee Ru‐Po , Grojean Emma M , Ly Austin , Hsu Mark , Heber David , Li Zhaoping
Published: November 2023

Preclinical studies have shown that the combination of quercetin (Q) and green tea (GT) extract enhanced the anti‐carcinogenic activity in a prostate cancer xenograft model, increased the bioavailability of GT polyphenols (P) and decreased P methylation compared to GT extract alone. In order to test the hypothesis in humans that Q in combination with GT extract would enhance bioavailability of GTP compared to GT extract consumption alone we enrolled 32 men diagnosed with prostate cancer and randomized them to receive 2 capsules of GT extract (240 mg) in combination with either 1 capsule of Q (400 mg) (GT+Q) or placebo (GT+PL) two times per day prior to prostatectomy. Blood and early morning spot urine were collected at baseline and at 3 weeks. Bioavailability of GTP and Q were tested at 3 weeks by sampling before and 2 hours after administration of GT and Q. Immediately after the surgery a small aliquot of prostate tissue was collected. All urine, blood and tissue samples were analyzed for GTP and Q by high performance liquid chromatography with coularray detection and mass‐spectrometry. Red blood cell (RBC) and prostate gene expression, protein expression and DNA and polyphenol methylation enzyme activity were determined. There was no liver toxicity in any subject. Q was increased 14‐fold, 12‐fold and 4.5‐fold in plasma, urine, and prostate tissue, respectively, in subjects given GT+Q by comparison to GT+PL. Plasma ECG was significantly increased in men consuming GT+Q compared to GT+PL consistent with an effect of Q on bioavailability of GT as previously observed in animals. Urinary excretion of EGC, 4'‐MeEGC and EC was significantly decreased in urine collected prior to surgery in participants consuming GT+Q compared to GT+PL. The percent of methylated EGC tended to increase from 36 to 79% in urine from men consuming GT+Q compared to GT+PL (p=0.137). ECG and/or EGCG were found in the prostate tissue of 31 out of 32 participants, but there was no difference in prostate GTP content between the GT+Q and GT+PL groups. There were also no epigenetic differences in prostate or RBC as determined by catechol‐methyltransferase and DNA‐methyltransferase gene and protein expression and enzyme activity. Further investigations are underway to determine whether the presence of quercetin in addition to GT polyphenols in the prostate modulated inflammation or oxidant stress in prostate tissue.

Support or Funding Information

NIH RO3CA171583
Characterizing the Impacts of Early Interventional Ketogenic Diet on Brain Vasculature, Energy Metabolism, and Cognition
Authors: Ma David , Wang Amy , Hoffmann Jared , Bakshi Vikas , Parikh Ishita , Guo Janet , Armstrong Rachel , Hartz Anika , Bauer Bjoern , Lin Ailing
Published: November 2023

The ketogenic diet (KD), or the low‐carbohydrate and high‐fat diet, has served as a therapeutic for medically intractable epilepsy for the past ninety years. Recent studies have shown the neurological benefits derived from ketone bodies. However, KD's impact on overall in vivo brain function remains largely unexplored. The aim of this study was to characterize the interaction between specialized nutrition and in vivo brain function. Subjects were age‐matched and gender‐matched young wild type mice models, divided into two groups based on diet: Western (control) diet or KD. We employed multimodal, non‐invasive neuroimaging (MRI/MRS) to determine in vivo brain cerebral blood flow and energy metabolites. We also assessed the animal's memory and learning ability with the Radial Arm Water Maze and the Novel Object Recognition Test. In addition, we performed western blots and BBB function analysis. Blood glucose, blood ketone bodies, and body weight were also measured. We found distinct patterns between Western and Ketogenic diet – the KD group exhibited significantly higher cerebral blood flow in the dorsal thalamus and the hypothalamus compared to the control. We observed significant modulation of the metabolites alanine and lactate, and identified a potential interaction with astrocytes. When examining the behavioral test results, there is indication that KD fortifies various memory and sensory functions, consist with our CBF data. Furthermore, KD mice exhibited significantly higher brain endothelial NOS and brain capillary P‐glycoprotein, as well as a significantly lower expression of the mechanistic target of Rapamycin. Our novel findings demonstrate KD produces noticeable shifts in brain vascular and metabolic function, while maintaining cognition in a young mice model. These results provide rationale for KD as a viable early interventional dietary measure.

Support or Funding Information

This research was supported by NIH grant K01AG040164.