bluuebunny/crossref_metadata_2025 huggingface dataset:
Endocrine response in administration of 5‐HT1A agonist associated with E2 replacement in ovariectomized rats.
Authors: FONSECA FABRICIA , Mecawi Andre , Araujo Iracema , Monteiro Lívia , Pereira Gislaine , Rodrigues José , Reis Luis
Published: June 2021

Series of studies support an estrogen modulation upon the serotonergic system activity. Thus, we examined the endocrine response in ovariectomized (OVX) rats submitted to oil or estrogen treatment (OVX‐E2, 80microg/kg, SC, 14 days) and to treatment with the 8‐OH‐DPAT(250microg/kg, 7 days) or vehicle (0.9% NaCl) during 7 days), in basal condition and after sodium depletion. We also investigated a possible correlation between serotonergic functional status and E2 replacement on the plasma levels of angiotensin I and II(ANG I, II), oxytocin (OT) and corticosterone (CORT) in basal condition and after sodium depletion. The ANG I and ANG II release and respective mechanisms seem to be depending of the condition of the animal or 8‐OH‐DPAT treatment. Chronic 8‐OH‐DPAT in basal condition increased the ANG I plasma concentration while the E2 seems to attenuate this response following sodium depletion. The OT and CORT plasma levels at basal condition were enhanced by association of the two treatments. E2 replacement, both at baseline and depletion conditions, increased OT and CORT plasma levels. Additionally we did not observe estrogen influence on plasma CORT concentrations following sodium depletion. Both serotonergic signaling as estrogen may be considered controlling systems of electrolyte homeostasis.
Loss of GABAergic Inhibition in Hypothalamus in Chronic Stress
Authors: Gao Yonggang , Zhou Jing‐Jing , Kosten Therese , Li De‐Pei
Published: June 2021

Chronic stress increases activity of hypothalamic‐pituitary‐adrenal (HPA) axis. CRH‐expressing neuron in the PVN is an essential component of HPA axis and plays a critical role in stress response. The activity of CRH neurons is regulated by excitatory and inhibitory GABAergic synaptic inputs. We hypothesized that chronic stress diminishes GABAergic inhibition, which contributes to hyperactivity of PVN‐CRH neurons. We identified PVN‐CRH neurons by expressing green fluorescent protein (GFP) driven by CRH promoter. Chronic unpredictable mild stress (CUMS) significantly increased plasma corticosterone levels. Blocking GABAA receptor significantly increased plasma corticosterone levels in unstressed rats. Furthermore, it significantly increased firing activity of PVN‐CRH neurons in unstressed rats but did not significant change in CUMS rats, while GABAA receptor significantly decreased firing activity of PVN‐CRH neurons in unstressed rats but slightly increased in CUMS rats. CUMS treatment significantly increased Na+‐K+‐2Cl‐ cotransporter (NKCC1) protein and mRNA levels while decreased K+‐2Cl‐ (KCC2) protein and mRNA levels in the PVN tissue. Immunocytochemical staining showed that they are expressed on CRH immunoreactivity positive PVN neurons. In addition, treatment with NKCC1 specific inhibitor decreased the basal firing activity of PVN‐CRH neurons in CUMS rats, but not in unstressed rats. Furthermore, it restored gabazine‐induced excitatory effect on PVN‐CRH neurons in CUMS rats. These data suggest that GABAergic inhibition in the PVN is diminished due to upregulation of NKCC1 in chronic stress.
Glycan Targeted Vaccines for The Induction of Immunity
Authors: van Kooyk Yvette , Fehres Cynthia , Horrevorts Sophie , Bruijns Sven , Kalay Hakan , Ambrosini Martino , Garcia Vallejo Juan , Unger Wendy
Published: June 2021

Dendritic cells (DC) are specialized in the recognition of antigen and play a pivotal role in the control of immunity and tolerance. For antigen recognition DC express several C‐type lectins, that function as innate receptors that facilitate antigen uptake and presentation. Many of these receptors also modify responses through signalling interference with TLR. We and others have shown that the C‐type lectins DC‐SIGN, MGL and Siglecs recognize specific glycan structures on pathogen related antigens but also on tumours. We have identified glycan structures that can either contribute to the tumour suppressive programming of the immune system, or oppositely activate the immune system.

By modifying antigens with specific glycans we can favour directional targeting of tumour antigens to human DC and Langerhans Cells (LC) in situ, in the skin, facilitating the induction of tumour specific CD4+ and CD8+ T cell responses and Th1 differentiation. Therapeutic vaccination of mice with glycan modified antigen show long term anti‐tumour immunity when tumour induced T regulatory cells are temporarily reduced. All together our work sheds new light on the contribution of glycans to control tumour immunity by either contributing to the induction of anti‐cancer immunity or tumour induced immune suppression.
Chronic Phenytoin Treatment Reduces Hypoxic Ventilatory Responses in Rats
Authors: Alcayaga Julio , Oyarce Maria Paz , Iturriaga Rodrigo
Published: June 2021

Arterial afferents provide sensory information that participates in the peripheral control of ventilation. The carotid body chemosensory afferents increase their frequency of discharge in response to hypoxia, increasing ventilatory frequency and volume. Acute phenytoin application, a persistent Na+ current blocker, reduces the basal afferent activity and hypoxia‐induced ventilatory responses. Because phenytoin is used as an antiepileptic drug, it is of interest to investigate the long‐term effects of phenytoin treatment on ventilatory control. Male Sprague Dawley rats (180‐200 g) were anesthetized with isoflurane, an osmotic pump (Alzet 2ML4) placed subdermally, the incisions closed and the animals received antibiotic and antiinflammatory. The osmotic pump delivered a volume of 2.59 µL/hr, with a phenytoin dose of 10 mg/day. Between one and four weeks after implantation, animals were placed in a whole body plethysmograph (Buxco, USA) and the ventilatory volume and frequency was measured at different inspiratory oxygen fractions (FIO2: 100, 50, 21, 15, 10, 7). The animals were sacrificed after four weeks by anesthetic overdose (pentobarbitone, 120 mg/kg). Phenytoin treatment had no effect on basal ventilation, but produced a marked reduction in the hypoxia induced ventilatory responses both in frequency and volume. Thus, minute ventilation was reduced at all hypoxic levels, with approximately a 40% reduction at 7% FIO2. Our results indicate that chronic phenytoin treatment reduces reflex ventilatory responses to hypoxia. It is suggested that a detrimental reduction of ventilatory chemosensory peripheral drive could result from chronic phenytoin treatment in epileptic patients.
Constant rules to guide nutrition guidelines
Authors: Thomas Diana , Heymsfield Steven
Published: June 2021

Purpose

Because of their simplicity, constant rules like the 3500 kcal rule, the 5% weight loss rule, and the three‐quarter fat free mass‐fat mass rule are being applied to offer nation‐wide weight loss advice, guide treatment, and inform experimental design. Despite their wide‐spread application, the accuracy of constant rules are rarely tested.

Methods

We tested the capacity of three different constant rules currently being applied to predict weight loss, determine effectiveness of weight loss treatment, and designate appropriate safe degree of fat‐free mass loss during weight loss against experimental data. The results were then compared to new models personalized to individual patient characteristics.

Results

Comparison of the 3500 kcal Rule to experimental data yielded inaccuracies of higher than 30 kg at three months even when subjects were placed on reasonable energy deficits. Similarly, prediction of long‐term weight loss success relying on only percent weight loss was not better at identifying true positives than random chance. Finally, we can show that loss of fat free mass during energy restriction is a non‐constant function of aging and gender. On the other hand, emerging dynamic and statistical models which incorporate continuous changes in physiology and consider covariates of age, height, gender, and baseline parameters, yield a substantially higher predictive accuracy on all fronts.

Conclusions

Constant rules have been historically applied in patient care because of their simplicity. However, inaccurate predictions can have severe implications contributing to reduced patient motivation or poor experimental design. We suggest the field move toward more physiologically sound and clinically tested models delivered through web‐based software.
Moderately High Protein Diets During Resistance/Aerobic Exercise Training Improve Body Composition Via Positive Changes in Adiposity But Not Lean Mass Accretion
Authors: Gordon Susannah , Kim Jung Eun , Weinheimer‐Haus Eileen , Campbell Wayne
Published: June 2021

Middle‐aged overweight/obese adults are faced with an environment of food abundance. This food excess challenges their ability to adjust eating behaviors and maintain body weight. We conducted a 36‐week diet supplementation and exercise training study where 35‐65 year‐old overweight and obese adults were exposed to a “controlled source” of potentially excess energy via 200 kcal supplements with varying amounts of maltodextrin and whey protein. The supplements were consumed 2x daily with breakfast and lunch along with their otherwise unrestricted, self‐chosen diets. Training included resistance (2 d/wk) and aerobic (1 d/wk) exercises. This retrospective data analysis assessed the effect of total protein intake (TPro) on changes in body weight and composition. 85 subjects were classified into 3 groups based on TPro: < 1.0 (low TPro, LP, n=29), 蠄 1.0 to < 1.2 (medium TPro, MP, n=24), and 蠅 1.2g•kg‐1•d‐1 (high TPro, HP, n=32). Weight was measured bi‐weekly and body composition (plethysmography) every 9 weeks. Over time, LP progressively gained weight and fat mass, while MP and HP maintained weight and lost fat mass. All three groups gained lean body mass (mainly during the first 9 weeks). These findings support that moderately high protein intake improves weight control and adiposity, but does not augment lean mass accretion among exercising overweight/obese adults who are challenged by potential excess energy intake. High protein intakes do not enhance these responses.

Support: US Whey Protein Res Consort; NIH T32AG025671; NIH UL1TR001108; Purdue Ingest Behav Res Ctr
Cranberry Extract Ameliorates Obesity‐Related Liver Inflammation via the Toll‐Like Receptor 4 Pathway
Authors: Glisan Shannon , Lambert Joshua
Published: June 2021

Obesity is commonly associated with an increase in hepatic triglycerides, known as nonalcoholic fatty liver disease (NAFLD). NAFLD may progress into nonalcoholic steatohepatitis (NASH), which includes an inflammatory component, and eventually cirrhosis. Polyphenols, such as those found in cranberries, have been shown to have antioxidant and anti‐inflammatory properties. The objective of this study was to determine the effect of cranberry extract (CBE) on NASH in an obese mouse model. C57bl/6J mice fed either 60%kcal from fat diet (HF, n=48) or 10%kcal from fat diet (LF, n=12). After 11 weeks, 24 mice from the HF group were switched to HF diet supplemented with 0.8% CBE. After an additional 10 weeks, mice were euthanized and markers of obesity and NAFLD were examined. Body weight, relative liver weight, and liver triglyceride concentrations were not affected by CBE supplementation. CBE supplementation did significantly decrease circulating alanine aminotransferase levels compared to the HF group, indicating a decrease in liver injury. Supplementation with CBE resulted in a decrease in hepatic toll‐like receptor 4 gene expression (63%) and hepatic nuclear factor (NF)‐κB gene expression (24%) compared to the HF group. Hepatic expression of tumor necrosis factor alpha, a pro‐inflammatory cytokine gene regulated by NF‐κB, and C‐C chemokine receptor 2 (CCR2), a receptor for monocyte chemoattractant protein‐1 was significantly decreased by CBE supplementation compared to HF mice. In conclusion, CBE may mitigate obesity‐related hepatic inflammation and liver damage; however, additional experiments are needed to better understand the underlying mechanisms of action.
Feasibility and Acceptability of Group Cell Phones to Promote Optimal Breastfeeding in a Women's Microcredit Program in Nigeria
Authors: Flax V , Ibrahim A , Negerie M , Yakubu D , Leatherman S , Bentley M
Published: June 2021

We previously reported on a breastfeeding promotion intervention that successfully increased optimal breastfeeding practices using cell phone messaging and group learning sessions integrated into a women's microcredit program in Nigeria. Using a novel approach, one cell phone was given to each group of 5‐7 microcredit clients. The cell phone holder was instructed to share the weekly breastfeeding voice and text messages with group members. The feasibility and acceptability of group cell phones was measured through structured exit interviews with microcredit clients (n=195) whose babies were born during the intervention (target group), in‐depth interviews with phone holders (n=8) and non‐phone holders (n=8), and focus group discussions (FGDs) (n=16) with other microcredit clients. Exit interviews revealed that only 6% of women in the target group were phone holders, but 64% said the group phone worked well. 58% lived within 10 min walking distance of the phone holder and 52% said messages were shared once a week. The target group was motivated to try the recommended practices because they trusted the information (58%) and had support from others (36%). In‐depth interviews and FGDs indicated that most phone holders were elected, non‐phone holders had positive feelings toward phone holders, and the group phone met their needs. A few issues with message sharing and connectivity were noted. In summary, use of group cell phones to promote breastfeeding among microcredit clients is feasible and acceptable, and can be part of an effective behavior change package. Funding: Alive & Thrive small grants program
Infection‐Specific Responses of Breastmilk Leukocytes
Authors: Hassiotou Foteini , Savigni Donna , Underhill Jenny , Moles Laura , Hartmann Peter , Geddes Donna
Published: June 2021

Breastmilk leukocytes comprise a small population of mature human milk when both the mother and infant are healthy, providing active immunity and immunocompetence in the infant and protecting the mammary gland from infection. In this study, we examined leukocyte‐subset responses to different infections of the breastfeeding dyad in mature breastmilk. Breastmilk leukocyte content was measured by flow cytometry using the pan‐immune cell marker CD45, and leukocyte subtypes were examined by co‐staining for CD14, CD3, CD4, CD8, CD56, CD19, and CD15. When both the mother and infant were healthy, breastmilk contained 0‐2% of leykocytes. However, when either the mother or the infant had an infection, breastmilk leukocyte content increased to up to 94% of total cells, with the greatest increase observed in cases of mastitis. During the latter, the majority of immune cells were activated T‐cells that responded to viral peptides if stimulated in vitro, with corresponding proliferation and cytokine secretion. Interestingly, leukocyte content returned to baseline levels (蠄2%) upon recovery from the infection. Similar responses were observed for immunoglobulins and lactoferrin. Different leukocyte subset profiles were recorded for different infections, with T‐cells differing the most between infections (P<0.004). These results demonstrate that breastmilk leukcyte content is dynamic, responding to the needs of the infant and reflecting the health status of the lactating breast. This is a new tool that can be potentially used to facilitate rapid diagnosis and treatment of conditions of the lactating breast.
Milk osteopontin may promote brain development via up‐regulating osteopontin expression in vivo
Authors: Jiang Rulan , Prell Christine , Lönnerdal Bo
Published: June 2021

Osteopontin (OPN) is a highly acidic phosphorylated glycoprotein that is involved in a wide range of biological processes, including biomineralization, immune modulatory functions, cell survival and proliferation. OPN is abundantly expressed in the brain and has been suggested to play a role in brain development. OPN is present in high concentration in human milk. In mice, OPN is also expressed in the mammary gland and secreted in milk. However, the functions of milk OPN remain unclear. In this study, wild‐type and OPN knock‐out mice were used to investigate the influence of milk OPN on brain development. To examine if dietary OPN can be absorbed and transferred to the brain, day 20 wild‐type mouse pups were fed I‐125 labeled OPN via oral gavage. Brain radioactivity was measured by gamma counter. After 30 min, 1h and 2h of incubation, radioactivity in the brain was detected at a stable level, around 1.7% of total OPN fed. Two groups, a control and an OPN‐deficient group, were then used to investigate the effects of dietary OPN on brain development. Wild‐type mouse pups were either fed by wild‐type dams (control) or by OPN knock‐out dams (OPN‐deficient group). Brain samples were collected every other day from D1 to D20. OPN mRNA expression was similar from D1 to D20 in both groups. Immunoblotting showed that peak OPN expression occurred at D4, D6 and D8 in both groups, but the control group expressed significantly higher concentrations of OPN compared with the OPN‐deficient group, suggesting post‐transcriptional regulation In addition, genes involved in brain development, including glial cell differentiation regulator and proteolipid protein were up‐regulated in the control group. In conclusion, milk OPN may play a role in brain development during early life. Supported by Mead Johnson Nutrition.
Selenium Binding Protein 1 Levels Predict Prostate Cancer Recurrence
Authors: Diamond Alan , Ansong Emmanuel , Deaton Ryan , Gann Peter
Published: June 2021

Selenium Binding Protein 1 (SBP1) is a non selenocysteine‐containing protein shown to be at lower levels in tumors and its lower levels are frequently predictive of a poor clinical outcome. Distinguishing indolent from aggressive prostate cancer (PCa) is a major challenge in disease management. Therefore, associations between SBP levels and 2 endpoints: tumor grade and disease recurrence following prostatectomy were investigated by duplex immunofluorescence imaging using a tissue microarray containing tissue from 202 PCa patients who experienced biochemical (PSA) recurrence after prostatectomy and 202 matched control patients whose cancer did not recur. Samples were matched by age, ethnicity, pathological stage and Gleason grade, and images were quantified using the Vectra® multispectral imaging system. Fluorescent labels were targeted for SBP1 and cytokeratins 8/18 to restrict scoring to tumor cells, and cell‐by‐cell quantification of SBP1 in the nucleus and cytoplasm was performed. Nuclear SBP1 levels and the nuclear to cytoplasm ratio were inversely associated with tumor grade using linear regression analysis. Following classification of samples into quartiles based on the SBP1 levels among controls, tumors in the lowest quartile were more than twice as likely to recur compared to those in any other quartile. Inducible ectopic SBP1 expression reduced the ability of HCT‐116 human tumor cells to grow in soft agar, a measure of transformation, without affecting proliferation. Cells expressing SBP1 also demonstrated a robust induction in the phosphorylation of the p53 tumor suppressor at serine‐15. These data indicate that loss of SBP1 may play a contributing role in PCa progression and monitoring its levels might be useful in distinguishing indolent from aggressive disease.
Dietary Fat Dose‐Dependently Increases α‐Tocopherol Bioaccessibility
Authors: Sapper Teryn , Mah Eunice , Chitchumroonchokchai Chureeporn , Failla Mark , Bruno Richard
Published: June 2021

Dietary intake of the lipophilic antioxidant α‐tocopherol (α‐T) is poor with a significant proportion of Americans having suboptimal α‐T status. Although its bioavailability is limited in the absence of co‐ingesting fat, the extent to which quantity and composition of fat affects α‐T bioaccessibility (BA), and its potential to be absorbed, is not well established. We hypothesized that bovine dairy fat would dose‐dependently increase α‐T BA and that dairy milk containing primarily saturated fat would increase BA to a greater extent than soy milk containing primarily unsaturated fat. In vitro digestion was used to examine the BA of α‐T (36 mg) added to soy milk (2.5 g fat) or dairy milk containing fat at <0.5 g (non‐fat milk), 2.5 g (low‐fat milk), 5 g (reduced‐fat milk), and 8 g (whole milk). α‐T recovery following simulated gastric through small intestinal digestion was 87‐94.5% with no difference (P>0.05) between dairy or soy milks. α‐T BA increased in a fat‐dependent manner (P<0.05) from 37.3±1.7% in non‐fat milk to 55.2±0.7% in low‐fat milk to 61.7±1.9% in reduced‐fat milk, and to 69.7±2.6% in whole milk. BA of α‐T in soy milk (53.5 ± 3%) and low‐fat milk with equivalent quantities of fat were not significantly different. Fat quantity (0‐8 g), regardless of dairy or soy milk, was highly correlated to α‐T BA (r = 0.92, P<0.05). Thus, dietary fat dose‐dependently increases α‐T BA independent of degree of lipid saturation and supports a need to define the optimal quantity of fat on α‐T bioavailability in clinical studies. Supported by Dairy Research Institute.
Neurodevelopment in Piglets is Influenced by Perinatal Choline Status as Evidenced by Magnetic Resonance Imaging
Authors: Mudd Austin , Getty Caitlyn , Dilger Ryan
Published: June 2021

Adequate choline supply during the perinatal period is critical for proper brain formation when robust neurogenesis and neuronal maturation are occurring. To examine the impact of perinatal choline on neurodevelopment, sows (n = 8) were fed a choline deficient (CD) or sufficient (CS) diet during the last 65 d of the 114 d gestation period. At 2 d of age, piglets from sows within each prenatal treatment group were stratified into postnatal treatment groups and provided either a CD or CS milk replacer, resulting in a 2 x 2 factorial arrangement (N = 32, n = 6‐9 piglets treatment). At 30 d of age, piglets underwent magnetic resonance imaging procedures to examine brain morphology and metabolite concentrations. Fractional anisotropy (FA) values, obtained from diffusion tensor imaging, revealed an interaction (P = 0.02) between prenatal and postnatal treatments in the right hippocampus. Pigs of a continuous perinatal CD status exhibited lower FA values than CS/CD and CD/CS piglets, highlighting the importance of perinatal choline supplementation for white matter development. Voxel‐based morphometry illustrated differences (P 蠄 0.01) between prenatal choline status in grey and white matter volumes of cortical, cerebellar, and hippocampal regions. Volumes of whole brain, cortical, and subcortical regions were greater (P 蠄 0.05) in prenatal CS compared with prenatal CD piglets. Hippocampal metabolites, assessed using single‐voxel spectroscopy, indicated a decrease (P < 0.01) in glycerophosphocholine + phosphocholine concentration in postnatal CD piglets and a decrease (P = 0.01) in glutamate + glutamine concentration in prenatal CD piglets. These results suggest that adequate dietary choline provided prenatally or postnatally, promotes neuronal growth and maturation.
Sex‐specific Effects of Prenatal and Post‐weaning High Fat Diet on Adult Mouse Offspring Skeletal Muscle Contraction
Authors: Jones Lisa , Markiewicz Filip , James Rob , Cagampang Felino , Rustighi Emiliano , Poore Kirsten , Newland Philip , Green Lucy
Published: June 2021

Maternal obesity is linked to altered offspring skeletal muscle structure and function. We tested the idea that prenatal (PRE) high fat (HF) diet would alter the deleterious effect of post‐weaning (POST) HF diet on offspring skeletal muscle contraction.

C57 mice were fed a control (C, 7% kcal) or HF (45% kcal) diet in PRE and/or POST periods (C/C, C/HF, HF/C, HF/HF). Male and female 30 wk offspring (n=7‐8/group) soleus and extensor digitorum longus (EDL) muscle peak force (PF) of isometric contraction was measured in response to several electrical stimulation frequencies.

In soleus and EDL, PF was greater in females than males. In female soleus (not EDL), POST HF alone markedly reduced PF (C/HF vs. C/C, P<0.01). In female soleus and EDL there was only a trend for a reduction in PF in HF/C and HF/HF groups (P=0.1). In male soleus and EDL, combined PRE and POST HF tended to reduce PF (HF/HF vs. C/C, P=0.1). In male soleus (not EDL), POST HF tended to reduce PF in PRE HF animals (HF/HF vs. HF/C, P=0.1), and PRE HF tended to reduce PF in POST HF animals (HFHF vs. C/HF, P=0.1).

Thus reduced adult skeletal muscle isometric contraction force by POST HF diet is modified by a prior PRE HF diet in a sex‐ and muscle bed‐specific manner. This could reflect developmental differences in myofibre type composition or lipid deposition, and has potential implications for muscle health in adult/ageing obese individuals.

Supported by The Gerald Kerkut Charitable Trust and Diabetes UK.
Evaluating the Discrimination of Sugar Content Thresholds in the Canadian Nutrient File Classification System
Authors: Bernstein Jodi , L'Abbe Mary
Published: June 2021

The objective of this study was to evaluate the discrimination of sugar thresholds in the Canadian Nutrient File Classification System (CNFCS). The CNFCS was developed and validated to evaluate the consistency of food and beverages with dietary guidance such as 'limit food and beverages high in sugar'. The CNFCS classifies products into one of four tiers separated by upper and lower thresholds for sugar, sodium, fat, and saturated fat content. Most choices should come from tiers 1&2, few from tier 3, and limited from tier 4. Unlike the other nutrients, sugar has no intake recommendations to base the CNFCS thresholds on. Assessing the discrimination of nutrient classification systems is integral as they categorize the overall healthiness of a food. Using the University of Toronto 2013 food database, the proportion of breakfast cereals (n=250) and yogurts (n=240) that met and/or exceeded the thresholds for sugar content was assessed and compared to the proportion categorized within each CNFCS tier. The majority of cereals (77%) and yogurts (85%) exceeded the lower threshold for sugar, but few (1% and 2%, repsectively) exceeded the upper thresholds. Despite surpassing the lower threshold for sugar, 64% of cereals and 55% of yogurts were classified into tier 1&2, based on other nutrient contents. In total, 86% and 65% fell into tier 1&2 and 3% and 8% into tier 4 for cereal and yogurt, respectively. These results demonstrate products with up to 19g of sugar for most food categories, or 28g for flavoured yogurt, could be categorized in the healthiest tiers. Few items fell in tier 4 or exceeded the upper sugar threshold, indicating CNFCS sugar criteria may be too lenient and hence provide little incentive for reformulation.

Funding: McHenry Chair (ML); CIHR PICDP & PHP (JB).
Endurance Capacity of Muscle‐specific Branched‐chain alpha‐Ketoacid Dehydrogenase (BCKDH) Kinase‐deficient Mice
Authors: Shimomura Yoshiharu , Xu Minjun , Ishikawa Takuya , Kadota Yoshihiro , Morioka Takashi , Toyoda Takanari , Kato Rina , Kondo Yusuke , Ota Miki , Morishita Yukako , Bariuan Jussiaea , Zhen Hongmin , Kitaura Yasuyuki
Published: June 2021

The main catabolic pathway of branched‐chain amino acids (BCAAs: leucine, isoleucine, and valine) is located in mitochondria. The rate‐limiting step of the catabolism in muscle tissues is catalyzed by BCKDH complex, which is located at the second step in the pathway. The activity of BCKDH complex is regulated by covalent modification, and BCKDH kinase (BDK) is responsible for inactivation of the complex by phosphorylation. In the present study, we produced muscle‐specific BDK knockout (BDK‐mKO) mice. The muscle BCAA concentrations were less than half in BDK‐mKO mice than in control mice. We assessed the endurance capacity of BDK‐mKO mice and control mice at 12 weeks of age (under untrained conditions) and at 21 weeks of age after 9 weeks training by measuring the distance of running to exhaustion using treadmill with 10% grade. The running speed in the test was initial 15 m/min for 4 min and then increased at the rate of 1 m/min every 4 min. The training was performed 5 days/week by running for 60 min/day at the speed of 15 m/min in the initial 4 weeks and 18 m/min in the following 5 weeks. The endurance capacity in control mice was 633 ± 109 m (mean ± SE, n=8) before training and 1,529 ± 165 m after training, and that in BDK‐mKO mice was 1,020 ± 45 m and 1,004 ± 110 m, respectively. These results indicate that BDK‐mKO mice, in contrast to control mice, did not adapt to the endurance training, suggesting that BCAAs play an important role in adaptation to exercise training.
Phenethyl Isothiocyanate (PEITC) from Cruciferous Vegetables Targets Human Cancer Stem‐Like Cells
Authors: Upadhyaya Bijaya , Liu Yi , Wang Dan , Knudsen David , Dey Moul
Published: June 2021

Scope

Cancer stem cells (CSC) are perceived as a key player in cancer chemo‐and‐radio‐therapy resistance, cancer recurrence, and metastasis. PEITC, a dietary bioactive compound, is pro‐apoptotic in various cancer cell lines and primary tumors but its effect on CSC is less known. Our objective was to examine the effects of PEITC on promoting apoptosis in human cervical HeLa CSC (hCSC) in vitro and on attenuation of tumorigenicity in hCSC‐transplanted mice.

Methods and Results

PEITC attenuated proliferation of sphere‐culture‐enriched (ANOVA, p蠄 0.001), aldehyde dehydrogenase (ALDH1)bright, CD44high⁄+/CD24low⁄–, Hoechst 33342‐excluded hCSC in a concentration‐ and time‐dependent manner. PEITC up‐regulated cleaved poly (ADP‐ribose) polymerase (p蠄 0.05) and induced death receptors, DR4 (p蠄 0.01) and DR5 (p蠄 0.001), of tumor necrotic factor‐related apoptosis‐inducing ligand signaling. In NOD/SCID mice, hCSC induced greater tumorogenicity, elevated serum human‐vascular endothelial growth factor A (ELISA), and lung metastasis (H&E staining) compared to HeLa cells, which were attenuated in 10 µM PEITC‐pretreated hCSC group. Ongoing experiments indicate that PEITC down‐regulates ALDH1 (p蠄 0.01) (flow cytometry) and specificity protein 1 (immunoblotting), a transcription factor regulating activity of several multi‐drug resistance genes.

Conclusion and Significance

PEITC attenuates cell proliferation and tumorigenicity by promoting apoptosis in CSC. This presents a novel opportunity for a dietary approach for preventing cancer recurrence and improving therapeutic outcomes in patients.

Research Support‐ NIH (R00AT4245) and SD‐AES (3AH360)
Use of the Automated Self‐administered 24‐hour Recall (ASA24) in the Real World
Authors: Subar Amy , Mittl Beth , Zimmerman Thea , Kirkpatrick Sharon , Schap TusaRebecca , Wilson Maggie , Potischman Nancy
Published: June 2021

Objective

To describe the use of NCI's ASA24 in real‐world studies.

Methods

Descriptive statistics were generated to summarize usage characteristics across studies using 2014 versions of ASA24.

Results

Since Feb. 2014, 319 studies had registered to use ASA24, 10% of which requested ASA24‐Kids. The percentages of studies using the following setup options were: 88% ‐ allowed unscheduled logins; 60% ‐ completion of recalls from midnight‐to‐midnight vs. past 24‐hrs; 73% ‐ allowed multiple vs. single logins; 68% ‐ completion time restricted to 24 vs. 32 hrs. The percentages of studies including optional questions were: 49% ‐ dietary supplements; 82% ‐ location of meals; 46% ‐whom meals were eaten with; 45% ‐ use of TV/computer/mobile phone during meals; 35% ‐ source of foods consumed. Nearly all recalls were completed after a single login; mean times to complete recalls with and without supplement questions was 32 and 23 minutes, respectively. Mean number of foods and beverages reported/day was 13. Mean and median number of participants/study was 509 and 72, respectively (range: 1‐100,000); mean and median number of recalls per participant was 8 and 3, respectively (range 1‐335). Most researchers (90%) were affiliated with academic institutions or government agencies. About 37% of studies had research funding; 14% were NIH‐funded. Two‐thirds of studies are using ASA24 for epidemiologic, intervention or clinical research.

Conclusion

ASA24 is being used by a diversity of researchers in a variety of ways to meet their research and teaching needs. Funding: NCI.
The Feasibility of Teacher and Parent Volunteers Collecting Digital Image Data of Children's Fruit and Vegetable consumption during School lunch
Authors: Amin Sarah , Stickle Timothy , Yon Bethany , Howard Alan , Eriksen Harley , Cochran Rebecca , Johnson Rachel
Published: June 2021

Increasing children's fruit and vegetable (FV) consumption is an aim of school nutrition programs. Digital imaging (DI) is less time and resource intensive than other dietary assessment tools for evaluating whether these efforts correspond with increased FV consumption or waste. However, it is impractical for a university‐based trained dietary assessment team (UDAT) to collect DI data nationally. The study objective was to compare the feasibility of DI data collection by the UDAT to a parent volunteer dietary assessment team (PDAT) and a teacher dietary assessment team (TDAT) at two northeast elementary schools (NES‐A and NES‐B, respectively) across 19 data collection days. Uniquely labeled lanyards were distributed to children as they entered the cafeteria to allow for the formation of a matched DI pair: pre‐image of the lunch tray as the child exited the lunch line (FV selection) and a post‐image of the tray before disposal (FV waste). Feasibility was defined as the number of DI pairs captured out of the number of lanyards distributed (LD). At NES‐A, the PDAT was less proficient at collecting DI pairs (74.1%, n=218 of 294 LD) than the UDAT (81.9%, n=262 of 320 LD; RR=0.91, 95% CI=0.83‐0.99, p<0.05). At NES‐B, the TDAT was better able to capture DI pairs (95.9%, n=257 of 268 LD) than the UDAT (91.3%, n=366 of 401 LD; RR=1.05, 95% CI=1.01‐1.09, p<0.05). In a school cafeteria setting, teacher volunteers may be more suitable for collecting DI dietary assessment data than parent volunteers and university researchers.
Customer utilization of small stores in rural Maryland and association with diet and psychosocial variables
Authors: Shapiro Myra , Welsh Claire , Dennis Donna , Campbell Elizabeth , Gittelsohn Joel
Published: June 2021

Small stores are a common food source for rural residents that have limited access to supermarkets. Scant data has been published about rural small stores customers, leaving their store utilization and food purchasing habits unknown. The objective of this study was to investigate the association between small store usage and spending, diet, and food‐related psychosocial variables. Customers (n=311) were interviewed in small stores in 7 rural Maryland counties regarding their store use, spending, and food‐related psychosocial variables; the NIH fruit and vegetable intake screener and the NCI percentage calories from fat screener were also used. Multiple linear regression was used to investigate relationships adjusting for age and sex. Customers who visited small stores more frequently in the previous 30 days spent less per visit (p<0.01). Fat intake was greater among customers who utilized small stores more often (p<0.01); fruit and vegetable intake was not related to store use. Customers who frequented small stores more often had more unhealthy behavioral intentions regarding food (p<0.01) and a greater self‐efficacy regarding food (p<0.01). There was no relationship between store use and food‐related knowledge. Interventions in small rural food stores should aim to reduce dietary fat while increasing access to healthy foods. To encourage customers to improve their diets, increasing self‐efficacy regarding healthy food choices is an important tool. Special attention should be paid to customers' intent to purchase and consume healthy food to ensure it does not hinder adoption of healthy habits. Small rural food stores warrant further investigation as a potential intervention venue.
My People are Dying of Ignorance: Changes in Exclusive Breastfeeding Knowledge, Benefits, Barriers, and Misconceptions in Family Members Involved in a Breastfeeding Promotion Campaign
Authors: Reinsma Kathryn , Bolima Nancy , Fonteh Florence , Montgomery Susanne
Published: June 2021

Background

Since 2003, WHO has recommended exclusive breastfeeding (EBF) for the first 6 months of life. In the NW Region of Cameroon only 34% of women EBF for six months. Previous research indicates that family members positively influence women's breastfeeding practices, but are often excluded from breastfeeding promotion campaigns.

Study Objective

To design an Entertainment Education (EE) audio program to test the hypothesis that EE improves EBF knowledge, benefits, and decreases misconceptions and barriers among family members of breastfeeding women.

Methods

Formative research consisting of 12 semi‐structured interviews and 6 focus group discussions informed the content of an EE audio program using grounded theory. Differences in EBF knowledge, benefits, barriers and misconceptions between and within the control group and the intervention group were evaluated using pre‐and post‐tests and analyzed with Wilcoxon Sign Test and Mann‐Whitney statistical tests.

Results

The audio program significantly improved overall EBF knowledge and benefits and decreased misconceptions within the intervention group (p <. 05).

Conclusion

This study provided insight into family member's EBF knowledge, benefits, barriers, and false misconceptions and proved that an EE audio program is an effective method of providing breastfeeding education to this population.
Relationships Between Fluid Intake, Breast Milk Volume, and Urine Volume in Lactating Women
Authors: McKenzie Amy , Perrier Erica , Jimenez Liliana , Armstrong Lawrence
Published: June 2021

The purpose of this investigation was to determine if a relationship existed between total fluid intake (TFI; volume of fluids consumed from drinking water and other beverages) and breast milk volume (BMvol) in lactating women, with follow up evaluations of urine volume (Uvol), urine concentration, and total fluid output (TFO; Uvol+BMvol).

Methods

Eighteen lactating women (3.3±0.7 weeks postpartum, 31±3y, 1.66±0.07m, 72.06±17.26kg) recorded TFI and BMvol (estimated by measuring body mass of newborn immediately before and after nursing at each feeding) for 24h. Women also collected their urine for measurement of 24h Uvol and osmolality (Uosm).

Results

No significant relationship existed between TFI and BMvol (r2=0.0235, p=0.544); however, there was a significant relationship between TFI and Uvol (r2=0.5006, p=0.001) and TFI and TFO (r2=0.4857, p=0.001). Those who did not meet the European Food Safety Authority reference value for TFI (2.16L/d) had higher 24h Uosm (621±60 vs. 445±68 mOsm·kg‐1, p=0.069) and lower 24h Uvol (1.646±0.243 vs 2.352±0.331 L, p=0.1046).

Conclusion

These findings demonstrate that in lactating women, there is no relationship between TFI and BMvol, indicating that BMvol is defended over a wide range of TFI (1.212‐3.998L). However, this defense of BMvol may come at the expense of lower Uvol and higher Uosm for the mother.

Supported by Danone Research.
Evo‐Devo and an Expanding Evolutionary Synthesis
Authors: Carroll Sean
Published: June 2021

Biologists have long sought to understand which genes and what kinds of changes in their sequences are responsible for the evolution of animal anatomy and morphological diversity. Advances in molecular and evolutionary developmental biology over the past thirty years have led to a general understanding that animal form evolves largely by altering the expression of functionally conserved proteins, and that such changes largely occur through mutations in the cis‐regulatory sequences of pleiotropic developmental regulatory loci and of the target genes within the vast networks they control.
CD36, fibroblasts and intestinal inflammation
Authors: Reaves Titus , Nillas Andrea , Anderson‐Thomas Megan
Published: June 2021

Crohn's Disease (CD) is characterized by dysregulated intestinal inflammation with an unknown etiology. Intestinal fibrosis is a major pathophysiological complication of CD with features that include an overproduction of collagen, excessive contraction of intestine tissue, and intestinal stricture. Fibroblasts are mesenchymal cells that produce the excess collagen resulting in the aforementioned fibrosis. We have shown that intestinal fibroblasts express a variety of immune receptors. A reduction in CD36 (membrane‐bound and scavenger receptor that is an indicator of inflammation and interacts with TLR‐2, 4, and 6) on the surface of fibroblast results in a statistically significant reduction in transmigration of polymorphonuclear leukocytes (PMN) toward such fibroblasts. Dextran sodium sulfate (DSS) was administered to CD36‐deficient mice and age‐matched controls; DSS typically results in intestinal inflammation that is similar to that observed in Crohn's Disease. Results indicated that mouse PMN were less vigorous in their capacity to migrate toward fibroblasts and there was less inflammation observed in both the large and small intestine of CD36‐deficient mice. Results suggest that CD36 may play a prominent role in the inflammatory responses of fibroblasts in the intestine and highlight CD36 as a potential target receptor in the treatment of aberrant mucosal inflammation, which leads to fibrosis.
Nuclear Localization of β‐catenin Associates EMT with Aggressive Vulvar Squamous Cell Carcinoma
Authors: Holthoff E , Jeffus S , Kelly T , Quick C , Post S
Published: June 2021

Nuclear localization of β‐catenin in epithelial cells characterizes the epithelial‐mesenchymal transition (EMT) and is associated with tumor progression in numerous malignancies. Factors that contribute to aggressive behavior in vulvar squamous cell carcinoma (vSCC) are poorly defined; however, studies have shown that vSCC with an infiltrative pattern of invasion are associated with worse outcomes compared to those with nested/pushing patterns of invasion. This study proposes that infiltrative vSCC may acquire increased aggressive behavior through EMT‐like changes reflected by nuclear localization of β‐catenin.

Immunostaining patterns of β‐catenin were analyzed in 30 cases of vSCC: 10 cases with infiltrative pattern of invasion, 10 with nested/pushing pattern of invasion, and 10 with a “mixed” invasive pattern. Stained tumor sections were analyzed for nuclear localization or membranous staining of β‐catenin. A staining pattern was defined as predominant when present in >75% of tumor cells.

Predominant nuclear localization of β‐catenin was observed in 8 (80%) of the infiltrative tumors, compared to only 3 (30%) of the pushing/nested tumors. In contrast, membranous staining predominated in 5 (50%) of the nested/pushing tumors compared to only 1 (10%) of the infiltrative tumors. Both membranous staining and nuclear localization of β‐catenin was observed in 7 (70%) of the mixed tumors. Importantly, in 6 of the 7 mixed tumors (85%), β‐catenin showed membranous staining in the pushing regions and nuclear localization in the infiltrative regions.

Our results show that nuclear localization of β‐catenin is highly associated with an infiltrative pattern of invasion in vSCC, suggesting that development of an epithelial‐mesenchymal transition drives the more aggressive behavior in this subset of tumors.
The Pathogenesis of Familial Dysautonomia is Explained by Abnormal Elp1‐dependent Retrograde Nerve Growth Factor (NGF) Signaling
Authors: Tourtellotte Warren
Published: June 2021

Familial Dysautonomia (FD; Riley‐Day Syndrome; HSAN3) is characterized by debilitating sensory and sympathetic neuropathy. It is caused by a germline mutation of the Elp1 gene that leads to mis‐splicing and loss of Elp1 protein primarily in sympathetic and sensory neurons. How Elp1 functions in the pathogenesis of FD in developing and adult disease‐vulnerable neurons is not known.

We generated conditional knockout mice with Elp1 ablated specifically in sympathetic neurons and found that they have increased sympathetic neuron apoptosis, target tissue innervation abnormalities and dysautonomia similar to humans with FD. Sympathetic neurons isolated from Elp1‐deficient mice showed no abnormalities in their survival when they were completely immersed in medium containing NGF, a neurotrophic factor that is required for their survival and differentiation. By contrast, when sympathetic neurons were grown in partitioned cultures with NGF present only in the distal axon compartment to emulate the way NGF is acquired from peripheral target tissues in vivo, Elp1‐deficient neurons were found to have profound abnormalities in survival and axon outgrowth compared to wild type neurons. Thus, retrograde NGF signaling in sympathetic neurons is dependent on Elp1, and abnormalities in retrograde signaling explain the poor sympathetic neuron survival, abnormal target tissue innervation and resulting dysautonomia in patients with FD. Future studies to define how Elp1 regulates NGF retrograde transport and signaling may make it possible to target new signaling pathways for treating FD and other autonomic neuropathies for which no effective therapies currently exist (Support: NIH NICHD and OD).
Team‐based Learning in Gross Anatomy Improves Academic Performance and Perceptions of Teamwork
Authors: Killins Anita , Huitt Tiffany
Published: June 2021

As healthcare becomes more interdisciplinary in patient care, it is essential for students to master group problem solving and develop an appreciation for team collaboration. Team‐based learning (TBL) is a student centered teaching method that fosters high‐level cognitive learning and collaborative group dynamics. The study objective was to determine the effect of TBL in a gross anatomy laboratory on academic performance, quality of learning in team interactions, and student perceptions of teamwork.

Subjects were Doctor of Physical Therapy students at the University of Central Arkansas from 2010‐2013. The comparison group (n=102) received a traditional cadaveric gross anatomy lab. The experimental group (n=109) received a TBL hybrid approach wherein students alternated between traditional cadaveric dissections and TBL learning modules. We compared examination scores, overall course grades and survey data between groups.

The experimental group scored significantly higher on 3 of 4 written exams (p=0.014, p=0.015, p=0.001) and the overall course average (p=0.048). All students had positive attitudes towards the TBL method. The experimental group significantly rated perceptions of collaborating with peers as both positive (p<0.000) and necessary (p<0.000) and felt problem solving in a group was effective for learning (p<0.000). Also, the experimental group experienced growth throughout the course, improving their perceptions of teamwork as necessary for student success (p<0.000).

Our results support the implementation of TBL in the gross anatomy lab as an effective method for improving academic performance and developing positive attitudes toward interprofessional communication and teamwork.