bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of fat sources on high fat induced changes in inflammation and lipid metabolism in juvenile pigs (822.3)
Authors: Jang Saebyeol , Molokin Aleksey , Urban Jeseph , Solano‐Aguilar Gloria
Published: June 2021

Effects of fat sources on high fat induced changes in inflammation and lipid metabolism in juvenile pigs Saebyeol Jang1, Aleksey Molokin1, Joseph F. Urban, Jr. 1, Gloria Solano‐Aguilar1: 1USDA, ARS, Beltsville Human Nutrition Research Center, Diet, Genomics, and Immunology Laboratory, Beltsville, MD 20705

The goal of this study was to examine effect of anhydrous milk fat (AMF) and palm oil (PO) on the high fat induced changes in inflammation and lipid metabolism in juvenile pigs. Four‐week‐old conventional crossbred pigs were fed with either a low 10.8% fat (LF) or high 39.5% fat (HF) diets including AMF or PO as their main fat source for 18 weeks . Data was analyzed using Two‐Way ANOVA to determine main effect of fat source (AMF vs PO) or fat level (low vs high fat). Serum lipid panel analysis showed that cholesterol, HDL, and LDL levels were increased in high fat groups. In liver, expression of inflammatory genes including TNFα, iNOS and TLR4, was increased by high fat diets and mRNA expression of MCP‐1 was decreased in LF‐AMF and HF‐AMF (main effect of fat source, p = 0.01). High fat diets also decreased expression of lipogenic genes SCD1 and FASN in liver whereas increased SCD expression in isolated peripheral adipocyte. The mRNA expression for IL‐1β, TNFα, iNOS, COX2, TLR4, IL‐6, MCP‐1, ICAM and CD40 were increased in the stromal cell fraction associated with visceral fat from pigs consuming high fat diets. No significant main effect of fat source was detected in visceral or peripheral fat stromal fraction. Interestingly, hippocampal gene expression of TLR4, TNFα and MCP‐1 and serum endotoxin was lower in AMF groups showing main effect of fat source. Overall, this study showed that diet containing AMF attenuated inflammatory parameters in serum, liver and, brain compared with diet of PO in juvenile pigs.

Grant Funding Source: Supported by DRI #1101
Effect of providing a micronutrient‐fortified corn/soy atole or milk powder on linear growth in young Guatemalan children (828.6)
Authors: Villanueva Lisa , de Ponce Silvia , Alfonso Victor , Reinhart Gregory
Published: June 2021

A community‐based feeding study was undertaken in Retalhuleu among 939 malnourished 6‐72 mo old children (HAZ <‐1). Children resided in 18 villages in an impoverished region of southwest Guatemala. Children were randomly assigned to two groups to compare effects of two nutritional interventions on growth and micronutrient status. Group A (n=667) children received 18.75 g of Chispuditos®, a corn/soy atole fortified with 21 vitamins and minerals (delivering 12.5 mg of iron and 9 mg of zinc per day). Group B (n=272) children received an equivalent portion by weight of lactose free milk (diets were isocaloric and isonitrogenous). At the conclusion of 12 mo, the average HAZ score of Group A children increased 12.0% from ‐2.76 to ‐2.43 (p<.01) and by 7.3%, from ‐2.87 to ‐2.66 (p<0.01) in Group B. Children from both dietary groups exhibited improvements in linear growth, with Group A having a greater increase in HAZ than Group B (p=0.01). Formative research conducted with the micronutrient‐fortified corn/soy atole revealed the product was well accepted by both primary caretakers and children. During the study time period Chispuditos was one‐third the cost of lactose free milk and provided a greater linear growth benefit. In conclusion, Chispuditos® is a culturally accepted, fortified atole that improves the health and nutritional status of children from 6‐72 mo of age in a more cost‐efficient manner compared to milk powder.

Grant Funding Source: The Mathile Institute for the Advancement of Human Nutrition®
Antioxidant and anti‐inflammatory activities of cyanidin‐3‐glucoside and cyanidin‐3‐rutinoside in hydrogen peroxide and lipopolysaccharide treated RAW264.7 cells (830.23)
Authors: Jung Hana , Kwak Ho‐Kyung , Hwang Keum Taek
Published: June 2021

Cyanidin‐3‐glucoside (C3G) and cyanidin‐3‐rutinoside (C3R) are two major anthocyanins found in blackberry and black raspberry. Antioxidant and anti‐inflammatory effects of C3G and C3R in RAW264.7 murine macrophage cells were determined. The tested anthocyanins (10 and 20 µg/mL) significantly reduced H2O2‐induced cytotoxicity. Following 4 h incubation with C3G or C3R, intracellular reactive oxygen species and DNA damage measured by Hoechst and comet assay significantly decreased in H2O2‐stimulated RAW264.7 cells. Cellular ferric reducing antioxidant power also significantly increased with C3G and C3R treatment. Nitric oxide produced in LPS‐stimulated RAW264.7 cells treated with C3G and C3R was reduced by 39.7 and 34.0%, respectively. LPS‐induced prostaglandin E2 productions were also significantly inhibited by C3G (52.6%) and C3R (59.5%). Protein expressions of iNOS and COX‐2 decreased in the cells treated with the anthocyanins. Further, the tested anthocyanins down‐regulated NF‐κB expression, and up‐regulated I‐κB expression in the LPS‐treated macrophages. Overall results indicate that C3G and C3R, potent antioxidants, may suppress inflammatory response via inhibition of NO and PGE2 releases and modulation of gene expression.
Estrogen compromises the facilitatory effect of chronic nicotine on adenosine A2B receptor/K+ channel‐mediated renal vasodilations (837.3)
Authors: El‐Mas Mahmoud , El‐Gowilly Sahar , Elsalakawy Lamia , El‐Gowelli Hanan
Published: June 2021

We previously showed that the renal vasodilatory action of the adenosine analog 5’‐N‐ethyl carboxamidoadenosine (NECA) in female rats is mediated via adenosine A2B receptors (A2BRs) and is facilitated by estrogen (E2). We tested the hypothesis that renal NECA vasodilation and its estrogen and A2BR specificities are altered by chronic nicotine. Cumulative NECA (1.6–50 nmol) or papaverine (1‐243 nmol) vasodilations were not affected by nicotine (1‐8 mg/kg/day i.p., 2 weeks) in isolated phenylephrine‐preconstricted perfused kidneys of sham preparations. NECA, but not papaverine, vasodilations were reduced in kidneys of ovariectomized (OVX) rats and restored to near‐sham values after E2 replacement. Further, nicotine increased NECA vasodilations in OVX preparations in contrast to no effect in sham or OVXE2 kidneys. The enhanced NECA responsiveness in nicotine‐treated OVX preparations was abolished after the infusion of alloxazine (A2BR antagonist) or BaCl2 plus glibenclamide (blockers of inward rectifier and ATP‐sensitive K+ channels, respectively). Moreover, vasodilations caused by minoxidil (K+‐channel opener) were increased by nicotine in OVX, but not sham or OVXE2, preparations and this increase was abolished after BaCl2/glibenclamide infusion. Together, estrogen compromises the enhancing effect of nicotine on renal vasodilations evoked via the activation of the A2BR/K+ channel cascade.

Grant Funding Source: Supported by STDF ID 502, Egypt
Afferent and efferent immunological pathways of the brain: anatomy, function and failure (84.1)
Authors: Carare Roxana
Published: June 2021

Immunological privilege appears to be the result of two major processes: (a) unique lymphatic drainage systems for the brain; and (b) highly regulated specialised passage of inflammatory cells through the blood‐brain barrier. Most organs of the body have well‐defined lymphatic vessels that carry extracellular fluid to regional lymph nodes. The brain has no such conventional lymphatics. Instead, it has perivascular pathways that drain interstitial fluid (ISF) from the brain parenchyma and cerebrospinal fluid (CSF) from the subarachnoid space to cervical lymph nodes. Solutes and ISF drain along narrow, ~100 nm‐thick basement membranes within the walls of cerebral capillaries and arteries to cervical lymph nodes. This pathway does not allow traffic of cells from the parenchyma to lymph nodes. CSF drains into blood through arachnoid villi and through channels in the cribriform plate of the ethmoid bone into nasal lymphatics, permitting cells to travel from CSF to cervical lymph nodes. Efferent pathways by which lymphocytes enter the brain are regulated by integrins on lymphocytes and selective receptors on endothelial cells. I will review: (a) the functional morphology of lymphatic drainage of the brain; (b) mechanisms involved in the pathways by which lymphocytes enter the brain; and (c) the failure of lymphatic drainage of the brain parenchyma in Alzheimer's disease.
Evidence for ETB receptor regulation of circadian control of sodium excretion (860.1)
Authors: Johnston Jermaine , Speed Joshua , Pollock David
Published: June 2021

Lack of renal ETB receptor function exacerbates circadian blood pressure variation in response to a high salt intake. Thus, we hypothesized that ET‐1 maintains normal circadian variation of sodium and water excretion through activation of the ETB receptor. To test this hypothesis, Sprague Dawley (SD) or ETB receptor deficient (sl/sl) rats were given a 900 μEq Na salt load by oral gavage at the beginning of either their active (dark) or inactive (light) cycle (7 pm or 7 am). Urine was collected in 12 hr increments and mean arterial pressure (MAP) measured by telemetry throughout the study. SD rats excreted the salt load during the initial 12 hrs whether given before their active (941 ± 88 baseline active period vs. 1548 ± 76 μEq Na/12 hr active period post gavage) or inactive cycle (362 ± 33 baseline vs. 944 ± 57 μEq Na/12 hr post gavage). Sl/sl rats also excreted the salt load when challenged during their active cycle (609 ± 25 baseline vs. 1119 ± 97 μEq Na/12 hr post gavage). However, sl/sl rats retained the salt load in the first 12 hr post gavage when given prior to the inactive period (364 ± 53 baseline vs. 464 ± 83 μEq Na/12 hr post gavage), but then excreted the salt load during the subsequent active period (684 ± 88 baseline vs. 1342 ± 121 μEq Na/12 hr, 24 hrs post gavage) and was associated with a significant elevation in MAP. These data demonstrate that the ETB receptor contributes to the circadian control of sodium excretion and blood pressure.

Grant Funding Source: Supported by NIH grant P01 HL095499
Long‐term high salt diet delays development of renal injury and inflammation in murine systemic lupus erythematosus (860.12)
Authors: Broome Hanna , Ryan Michael
Published: June 2021

Systemic lupus erythematosus (SLE) is an autoimmune disorder with prevalent renal disease, for reasons that continue to be elucidated. Recent evidence suggests that dietary salt may be an important environmental factor that promotes some autoimmune diseases. Therefore, we hypothesized that a long‐term high salt diet would accelerate the progression of renal disease during SLE. In order to test this, an established experimental model of SLE (female NZBWF1 mice) was fed a standard (0.4% NaCl) or high salt (4% NaCl) diet starting at 10 weeks of age. Sodium intake (in meq/day) was measured three times throughout the study and was significantly greater in high salt fed animals at 16 weeks of age (2.9±0.6 vs. 0.5±0.06 , p=0.01), 23 weeks (4.8±0.8 vs. 0.6±0.06, p=0.004) and 29 weeks (5.7±0.6 vs. 0.6±0.05, p=0.0007). Urinary albumin was monitored monthly until 30 weeks then weekly by dipstick assay as a marker of renal injury until 34 weeks of age at which time the mice were euthanized and the kidneys were harvested. Renal cortex enriched mRNA was analyzed by qRT‐PCR for inflammatory markers. 80% of animals (n=5) on a normal diet developed albuminuria (蠅 100 mg/dL), whereas none (0%, n=5) of the high salt fed animals developed albuminuria. Renal cortical interleukin‐2 (IL‐2), a cytokine important for T cell differentiation, expression was significantly lower in high salt fed animals (0.26±0.14 relative to normal chow fed mice, p=0.03). These data suggest that, contrary to the original hypothesis, a long‐term high salt diet may protect against the renal injury associated with SLE, possibly through alterations in IL‐2 mediated T cell differentiation. Furthermore, the data advance our overall understanding of how dietary salt may impact different autoimmune disorders.

Grant Funding Source: Supported by T32HL105324, 12GRNT12060203, HL085907, and HL051971
Right ventricular function following left myocardial infarction in rats with and without heart failure (864.6)
Authors: Fernandes Aurélia , Moura Viviane , Siman Fabiana , Ribeiro Júnior Rogério , Dias Fernanda , Jorge Zoghaib , Vassallo Paula , Vassallo Dalton , Stefanon Ivanita
Published: June 2021

It has been proposed that right ventricular function is an independent predictor of mortality and development of heart failure in patients with known left ventricle dysfunction following myocardial infarction (MI). This study was conducted to assess right ventricle (RV) contractility 8 weeks following MI in rats presenting a moderate to large infarct area. Myocardial infarction was induced in male Wistar rats to create transmural infarctions involving 40‐60% of the left ventricle surface. Infarct rats were divided in two groups: those that presented classical signs of heart failure (HF group) and those that did not (INF group), and these two groups were used to study isometric contraction in isolated RV strips. Inotropic responses in the RV strips were preserved in the INF group but were reduced in the HF group (3.75 mM Ca2+ treatment: Sham=163±18; INF=148±19; HF=68±11 g/mg*; *p<0.05); (5x10‐5 M Isoproterenol treatment: Sham=151±15, INF=134±17, HF=52±7 g/mg*; *p<0.05). The infarcted group without signs of HF showed preserved contractility in RV strips in response to post‐rest potentiation, increases in extracellular Ca2+ and isoproterenol treatment. Indeed, SERCA‐2a protein expression was increased in the INF group but not in the HF group. In conclusion, increased SERCA‐2a protein expression may play a role in the preservation of RV function post‐MI. Therefore, therapeutic strategies that attempt to increase SERCA‐2a protein expression levels may be useful for the treatment of HF.

Grant Funding Source: Supported by FAPES and CNPq
Role of microRNAs in malignant heart diseases (868.5)
Authors: Yang Baofeng , Shan Hongli , Chu Wenfeng , Pan Zhenwei , Zhang Yong , Lu Yanjie
Published: June 2021

Deregulated myocardial fibrosis is a major cause of cardiac dysfunction. The focus of this investigation was to elucidate the importance of miRNAs in the control of pathophysiological processes following cardiac fibrosis. Gain of function and loss of function approaches in a specific miRNA transgenic mouse models were employed in the present study. First, we found that the level of the miRNA is aberrantly upregulated in the border zone of infarcted heart of wild type (WT) mouse following cardiac fibrosis by occlusion of the left coronary artery (LCA). Moreover, myocardial interstitial fibrosis has been dramatically developed in the genetic mouse model with cardiac‐specific overexpression of the miRNA. This phenomena in cardiac fibrosis followed by dilated cardiomyopathy and subsequent cardiac failure. In contrast, extent of fibrosis is significantly reduced by the genetic mouse model with cardiac‐specific knockdown of the miRNA by occlusion of LCA and elimination of the miRNA by its antisense oligoribonucleotides (AMO) in cultured cardiac fibroblasts significantly also results in reduction of secretion of collagen. In terms of its effect mechanisms, the miRNA directly targets at transforming growth factor‐beta receptor (TGFBR), which acts as an inhibitor of TGF‐β signals, subsequently promotes the formation of type I ‐ type II receptor complex and activities TGF‐β signaling pathway. Upregulation of TGF‐β signaling by the miRNA targeting TGFBR in turn induces secretion of collagen in fibroblast and downregulation of TGFBR is also abolished by the AMO in cultured cardiac fibroblasts. Our finding indicates that miRNA can function as a potent regulator of myocardial interstitial fibrosis and represents a potential therapeutic target for cardiac fibrosis in general.
Therapeutic effects and mechanisms of paeonol on cigarette smoke‐induced lung inflammation (869.13)
Authors: Liu Meng‐Han , Lee Hung‐Fu , Lee Tzong‐Shyuan , Kou Yu Ru
Published: June 2021

Cigarette smoke (CS) is known to cause persistent lung inflammation mainly regulated by redox‐sensitive pathways. Indeed, several investigations from our laboratory reveal that CS can sequentially increased intracellular ROS, activated AMP‐activated protein kinase (AMPK), mitogen‐activated protein kinases (MAPKs) and NF‐κB, increased IL‐8 production in lung cells and finally produced lung inflammation,. Thus, therapeutic targeting of oxidative stress with antioxidants is recognized to be beneficial in the treatment of CS‐induced lung inflammation COPD. Paeonol, a major component of Mu Dan Pi (Paeonia moutan), has been suggested to be a potential antioxidant and anti‐inflammatory drug against diseases in several organs. However, whether paeonol may decrease the CS‐induced oxidative stress and lung inflammation is unknown and what therapeutic mechanisms of this benefit are still unclear. In this study, we investigated the therapeutic effects and mechanisms of on CS‐induced lung inflammation. Using human bronchial epithelial cells (HBECs) received challenge by cigarette smoke extract (CSE) demonstrate the suppressive effects of paeonol on the CSE‐induced increase in intracellular ROS, activation of the AMPK/MAPKs/NF‐κB signaling pathway, and the subsequent induction of IL‐8 chemokine. Using a murine model chronic CS exposure for 4 weeks increased macrophage inflammatory protein 2, and induced lung inflammation. These CS‐induced events were alleviated by treatment with paeonol. Our results suggest a novel role for paeonol in suppressing the CSE‐induced IL‐8 in vitro via its antioxidant function and by inhibiting both ROS‐sensitive AMPK/MAPK signaling pathway and the downstream transcriptional factors NF‐κB and in improving lung inflammation induced by chronic CS exposure in vivo.
The role of arginine vasopressin and oxytocin as immunomodulatory hormones on experimental autoimmune encephalomyelitis (876.1)
Authors: Verdugo Meza Andrea , Carrasco Esparza Norma , Ramírez Esquivel Ubaldo , Viñuela Berni Verónica , Organista Esparza Alejandro , Berczi Istvan , Kovacs Kalman , Quintanar Stephano Andrés
Published: June 2021

Arginine vasopressin (AVP) and oxytocin (OXT) are pleiotropic hormones released from the neurohypophysis. We demonstrated that neurointermediate pituitary lobectomy (NIL) in rats significantly protects against experimental autoimmune encephalomyelitis (EAE), a demyelinating T cell‐mediated disease of the central nervous system (CNS), whereas administration of desmopressin (DP, an AVP synthetic analog), restores the susceptibility of the NIL animals to EAE. Since immune cells have AVP and OXT receptors, we investigated whether OXT beside AVP, has also an immunomodulatory role on EAE. A comparative study of the effects of NIL, NIL+ DP and NIL+ carbetocin (CBT, a synthetic OXT analog) on clinical symptoms of EAE and on the number of CNS infiltrating lymphocytes were assessed. Animals were immunized for EAE one week after starting the administration of DP and CBT. EAE clinical signs were evaluated daily and CNS lymphocytes were separated with Percoll gradient. In NIL rats DP restored susceptibility to EAE and increased lymphocytic infiltration in the CNS while CBT resulted in very mild EAE symptoms and no changes in CNS lymphocytes. In conclusion, AVP has more powerful immunomodulatory effects than OXT.
Murine airway slowly‐adapting receptor responses to lung inflation: a role for serotonin? (879.19)
Authors: Domnik Nicolle , Vincent Sandra , Cutz Ernest , Fisher John
Published: June 2021

The mechanical phenotype of murine airway slowly‐adapting receptors (SARs), which are mechanosensitive afferents physiologically characterized by maintained discharge to sustained transmural pressure, is only partially defined. Adriaensen et al. suggest that densely‐innervated pulmonary neuroepithelial bodies (NEBs) are a morphological equivalent of SARs (Adriaensen et al. 2006, J Appl Physiol). Since NEBs release serotonin (5‐HT) in response to stretch in vitro (Pan et al. 2006, Am J Physiol Lung Cell Mol Biol), we characterized the mechanosensitivity of SARs during quasi‐static inflation to 20 cmH2O in male C57BL/6 mice and tested the impact of 5‐HT3 receptor blockade (i.v. tropisetron, Trop; 4.5mg/kg). SAR (n=8) discharge frequency (f; impulses/s) mechanosensitivity was proportional to tracheal pressure. Trop caused a small (3‐8%) but significant decrease in f vs. control (P=0.005; 2‐way ANOVA‐RM; mean±SEM (imp/s): 5cmH2O ‐ con: 91±13, Trop: 87±13; 10cmH2O ‐ con: 153±25, Trop: 143±22; 15cmH2O ‐ con: 192±29, Trop: 180±28; 20cmH2O ‐ con: 176±14, Trop: 164±15). It remains to be seen whether such a modest reduction in SAR discharge has a physiologic impact on the reflex effects of SAR medullary input.

Grant Funding Source: Supported by: CIHR (JTF; MOP81211) & NSERC PostGrad Scholarship (NJD)
Profound metabolic depression in the hypoxia‐tolerant naked mole rat (879.2)
Authors: Pamenter Matthew , Dzal Yvonne , Milsom William
Published: June 2021

Naked mole rats (NMRs) are among the most hypoxia‐tolerant mammals identified. Metabolic depression is a common response to hypoxia and is associated with lowered body temperature and a reduced state of activity (e.g. torpor). Contrary to this, NMRs maintain body temperature and remain active during prolonged exposure to 3% O2. The physiological adaptations that enable NMRs to remain active and thrive in hypoxia are poorly understood and we asked whether hypoxia‐induced changes to metabolic, ventilatory, or cardiac homeostasis contribute to this tolerance. To examine this question we exposed NMRs to progressive hypoxic epochs of 21, 9, 7, 5, and 3% O2 and measured metabolic rate, physical activity, ventilation, cardiac work, and body temperature. We found that NMRs exhibited profound metabolic depression (> 85%) at hypoxic levels < 7% O2, while maintaining body temperature and physical activity. Ventilation and heart rate increased with hypoxia but at a lower O2 levels than in hypoxia‐intolerant animals. Metabolic depression was reversed by systemic injection of the adenosine receptor antagonist aminotheophylline. Following injection, exposure to 7% O2 became lethal to animals that previously tolerated 3% O2 without detriment. We conclude that the hypoxia‐tolerance of NMRs is dependent upon robust metabolic depression, which is mediated by a mechanism involving adenosine receptor activation.

Grant Funding Source: NSERC
Influence of exercise mode on maternal and fetal health outcomes (886.3)
Authors: Moyer Carmen , May Linda
Published: June 2021

Current research is exploring the health benefits of exercise in pregnancy. The purpose of this study was to compare the effects of aerobic and circuit training throughout pregnancy on maternal and fetal adaptations. We hypothesized that there would be 1) improvements in maternal resting HR and body composition; 2) differences in fetal HR, HR variability, and heart measures at 34 weeks; and 3) no differences in fetal anatomical heart measurements or birth measures between groups. Participants in one of three protocols completed three 45 minute sessions weekly from 13 weeks gestation to delivery. We measured maternal resting HR at each exercise session, maternal body composition monthly, and 34 week fetal heart measures. Statistical analyses included t‐tests and multiple ANOVAs. There is a significant difference in RHR between groups for OW/OB women, but no significant difference in body composition between exercising groups. There are differences in fetal HR, HR variability, and stroke volume between exercising and control groups, but no differences in fetal anatomical measures between groups. These data suggest that there is a similar fetal heart response between aerobic and circuit exercising groups that differed from the control group. Fetal cardiac autonomic benefits are observed when controlling for frequency, intensity, and time, regardless of the exercise protocol, compared to not exercising.
A computational‐experimental protocol for investigating the quantity and anatomical location of pre‐glomerular oxygen shunting (890.13)
Authors: Kar Saptarshi , Gardiner Bruce , Smith David , Evans Roger , Ngo Jennifer
Published: June 2021

Determination of the magnitude and location of pre‐glomerular oxygen shunting is critical in understanding its role in renal hypoxia. Unfortunately, experimental studies have been unable to quantify the magnitude and location of pre‐glomerular oxygen shunting. Computational models could theoretically fill this void by providing quantitative information related to renal oxygen shunting. However, their use is limited by uncertainty in model parameters. In this study we show the benefits of integrating oxygen pulse experiments with computational models of oxygen transport to quantitatively analyze oxygen shunting. The method involves injecting an oxygen pulse of known shape into the renal artery and monitoring the oxygen concentration profile at the renal vein exit. The shape and timing of the venous oxygen elution curve indicates the magnitude and location of oxygen shunting along the renal vasculature. Here, we developed a transient computational model for oxygen transport in pre‐glomerular vasculature simulating the above experimental protocol. The role of shunting on renal vascular oxygen transport is investigated by varying the shape of the input oxygen pulse in the renal artery and recording the timing and shapes of the corresponding exit pulses in the renal vein. The oxygen shunting parameters in our model can subsequently be fine‐tuned to match the pulse shapes from experimental data. The fine‐tuned model can then provide definitive answers about the magnitude and location of oxygen shunting.

Grant Funding Source: NHMRC Australia Grant # 384101 and 606601
The renal Na+‐driven Cl‐HCO3‐ exchanger SLC4A8 is important for maintaining sodium balance (892.36)
Authors: Jayat Maximilien , Sinning Anne , Gerth Melanie , Calderon Carolina , Cornière Nicolas , Baron Stéphanie , Hadchouel Juliette , Eladari Dominique , Hubner Christian , Chambrey Régine
Published: June 2021

We recently described a novel renal NaCl transport system expressed in intercalated cells of the cortical collecting duct (CCD) that results from the parallel action of the Cl‐/HCO3‐ exchanger pendrin and the Na+‐driven Cl‐/HCO3‐ exchanger (NDCBE/ Slc4a8). Even though a role for pendrin in maintaining Na+ balance, normal vascular volume and blood pressure is well documented, there is no direct evidence for such a role for NDCBE yet. We first demonstrated that NDCBE protein was upregulated by primary or secondary hyperaldosteronism, two situations known to increase renal NaCl absorption. Ndcbe deficient mice exhibited hallmarks of vascular dehydration as evident from increased renin mRNA expression and plasma renin activity and developed marked secondary hyperaldosteronism when subjected to NaCl restriction. Systolic blood pressure was lower in Ndcbe deficient mice fed either a normal or NaCl‐free diet and was normalized with a high salt diet, indicating that the lower blood pressure in Ndcbe deficient mice is the consequence of a decreased vascular volume. Since Ndcbe disruption was compensated by an increase in NCC expression and activity, we generated double Ncc/Ndcbe knock‐out mice. Volume depletion in these mice was more pronounced although expression of other major renal Na+ transporters was markedly increased. Taken together, these results demonstrate that NDCBE is critical in maintaining sodium balance, normal vascular volume and blood pressure.
Differential effects of chemotherapeutic agents used to treat multiple myeloma on viability of colon cancer and colon cancer stem‐like cell lines (899.5)
Authors: Lenzi Kerry , Velez Hannah , Knabb Maureen
Published: June 2021

Cancer stem cell (CSC) proliferation is one of the underlying causes for recurrence of malignancy. Chemotherapeutic agents used to treat multiple myeloma following stem cell transplant (SCT), such as lenalidomide (L) and bortezomib (B), are known to increase the risk of secondary cancers. Accordingly, the cytotoxic effects of L and B were investigated in human colon cancer HCT‐116 (C) and 5‐Fluorouracil/oxaliplatin (FOLFOX)‐resistant HCT cells. These FOLFOX‐resistant (R) cells are known to express CSC markers. Cells were cultured in DMEM with 10% FBS and cell growth inhibition was assessed using Dojindo CCK‐8 cell proliferation assay in 96‐well plates. In C cells, FOLFOX inhibited growth by 50% at a dose of 50 µM 5‐Fluorouracil + 1.25 µM oxaliplatin. A 500‐fold dose was required to inhibit growth by 50% in R cells. C cells were highly sensitive to B with an LD50 of 12 nM, but R cells remained viable at a 100 µM dose. L failed to induce cytotoxic effects in R and C cell lines. A higher sensitivity of colon cancer cells to bortezomib indicates that this would be the more efficacious maintenance therapy following stem cell transplant in MM patients. However, as evidenced by CSC resistance to bortezomib and lenalidomide, patients treated with these chemotherapeutic agents may be at risk for increased incidence of secondary cancer growth following SCT.

Grant Funding Source: supported by WCU CASSDA award
Iron deficient toxic milk leads to the mask phenotype in hephaestin knockout mice (907.4)
Authors: Page Kathryn , Lachenauer Erica , Luong Julie , Han Chenchen , Irimagawa Hiroko , He Yuanchi , Killilea David , Frazer David , Anderson Gregory , Fuqua Brie , Vulpe Chris
Published: June 2021

Iron is an essential element that is required for many cellular functions. Multiple symptoms of iron deficiency have been reported in humans, including anemia, hair loss, stunted growth, and reduced oxygen transport. Mammalian iron intake is mediated in two different ways, across the placenta during fetal growth, and through the intestine during postnatal growth and development. We previously identified hephaestin (Hp), a copper‐iron ferroxidase, which plays a key role in intestinal iron transport. Hephaestin oxidizes iron and works in conjunction with ferroportin to facilitate iron export from enterocytes and its efficient binding to plasma transferrin. Null mutations in the Hp gene cause an accumulation of intracellular iron, specifically in the intestine. During analysis of the Hp null mice, we discovered that neonates born to Hp knockout (Hp KO) dams developed truncal hair loss (the “mask” hair loss phenotype) that resolved after weaning. The same phenotype was also observed in pups born to mothers with intestine‐specific knockout of Hp (even WT (floxed) pups born to these mothers), suggesting that lack of maternal Hp in the intestine alone is responsible for the mask phenotype. In order to determine the stage (ante and/or post natal) at which lack of maternal Hp leads to pup hair loss, we performed cross‐fostering studies. Wild‐type pups fed by Hp KO dams presented with the “mask” phenotype, whereas, all pups fed by the WT dams had normal hair growth. Histology of the affected skin of pups fed by Hp KO dams showed disorientation and keratinization of the follicles, follicular infundibular plugging and some tortuosity to the hair canals. Our findings suggest the involvement of a “toxic milk” phenotype involving low iron levels in the milk causing this phenotype.

Grant Funding Source: R01 GM083198‐01A1
Stretchy nerves withstand deformation associated with lunge feeding in rorqual whales (918.21)
Authors: Vogl A. , Lillie Margo , Piscitelli Marina , Shadwick Robert
Published: June 2021

Rorqual whales have evolved a remarkable feeding strategy that involves rapidly gulping enormous quantities of prey‐laden water and then more slowly separating the prey from the water by filtering the material through baleen plates suspended from the roof of the oral cavity. During the initial engulfment, the tongue inverts into a ventral pouch and balloons backwards as far as the umbilicus. Simultaneously muscles in the floor of the oral cavity, and the overlying ventral groove blubber, dramatically expand to accommodate the engulfed water that can reach a volume exceeding that of the whale itself. We have found that large nerves in the tongue and in the floor of the oral cavity of adult fin whales can stretch or extend up to twice their initial lengths. This contrasts with intercostal nerves that are virtually inextensible. Histological analysis demonstrates that nerves in the tongue and floor of the oral cavity are surrounded by a very thick connective tissue coating consisting of elastin and collagen, and that the central nerve bundle itself is folded or coiled into a small central core ‐ not unlike a coiled string inside a much thicker bunge‐cord. Intercostal nerves are not folded and have much thinner surrounding walls. Stretchy nerves are yet another extreme morphological adaptation to lunge feeding in rorqual whales.

Grant Funding Source: Supported by NSERC Discovery Grants
Localization of the recurrent laryngeal nerve using ultrasound (923.5)
Authors: Snosek Michael , Margarit Georgiana , Burns Danny , Hage Robert , Tubbs Shane , Loukas Marios
Published: June 2021

INTRODUCTION: The anatomical course of the recurrent laryngeal nerve (RLN) deep to the thyroid gland predisposes it to iatrogenic damage during thyroid surgery. The course of the nerve and its anatomical variations are well documented in the literature. The aim of this study was to localize the RLN in relation to key surgical landmarks with the aid of non‐invasive modalities such as ultrasound. METHODS: We examined 50 adult healthy individuals with the aid of Logiq E ultrasound unit using a 12Hz linear transducer. In addition, 20 cadaveric specimens were examined with the aid of ultrasound and then dissected to identify the RLN. RESULTS: The RLN was identified in 10 individuals (20%) as a linear hypoechoic structure in the tracheoesophageal groove. The RLN was found in 6 cadaveric specimens. Despite the fact that the RLN itself was not identified in the majority of the necks, the ultrasound can identify the plane between the thyroid gland and the esophagus, where the RLN can most commonly be found. CONCLUSIONS: This study introduces a new, non‐invasive technique for localizing the RLN and the space between the thyroid and the esophagus. This technique might be of benefit for presurgical localization of the RLN or ultrasound‐guided needle procedures.
Identification of the most immunogenic regions in vaccine candidate P6 (948.7)
Authors: Bettinger John , Newman Emily , Mangan Anthony , Pichichero Michael , Michel Lea
Published: June 2021

P6 is a leading protein vaccine candidate for protection against Nontypable Haemophilus influenzae (NTHi) infection. NTHi bacteria are a cause of middle ear infections, sinusitis, pneumonia, and chronic bronchitis. Vaccines that contain only the most immunogenic parts of the protein, often including the antibody binding sites or “epitopes,” can reduce production costs and can sometimes enhance effectiveness. Thus, identifying the most immunogenic part of P6 might be highly attractive to pharmaceutical companies who would like to use P6, or part of P6, in their next vaccine against NTHi. In this study, the P6 epitopes against monoclonal antibodies 4G4 and 7F3 were partially identified. P6 mutant T42R (threonine to arginine at residue 42) was prepared using recombinant DNA technologies, and mutated DNA was transformed into Escherichia coli cells for efficient overexpression. An Enzyme‐Linked Immunosorbent Assay (ELISA) showed decreased binding (compared to wild‐type P6) of the P6 T42R mutant for both the 4G4 and 7F3 antibodies, suggesting that 7F3 and 4G4 interact with residue 42. Nuclear Magnetic Resonance (NMR) spectroscopy was employed to assess structure changes in P6 due to the mutations.

Grant Funding Source: This study was supported by NIH NIDCD RO1 08671 (to MEP) and the Rochester Institute of Technology.
Comparison of biological activities of the leaf and root extracts of Smilax china L. (959.11)
Authors: Kang Yun Hwan , Kim Dae Jung , Kim Tae Woo , Kim Kyoung Kon , Choe Myeon
Published: June 2021

This study was carried out to compare the biological activities of leaf and root water extracts of Smilax china L. (SC) by measuring the total polyphenol and flavonoid contents, anti‐oxidant activity, inhibitory effect on α‐glucosidase, and anti‐inflammatory gene expression. The total polyphenol and flavonoid contents of SC leaf (SCLE) and root (SCRE) water extracts were 127.93 mg GAE/g and 39.50 mg GAE/g and 41.99 mg QE/g and 1.25 mg QE/g, respectively. The anti‐oxidative activities of SCLE and SCRE were measured using 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay and reducing power assay. Both SCLE and SCRE scavenged radicals in a concentration‐dependent manner, and SCLE showed stronger radical scavenging activity and reducing power than those by SCRE; however, the activities of both SCLE and SCRE were lower than that of ascorbic acid. Compared to the anti‐diabetic drug acarbose, which was used as a positive control, in SCLE and SCRE, the α‐glucosidase inhibition activities was low, nevertheless, the activity of SCLE was 3.7 fold higher than that of SCRE. Finally, the SCLE significantly decreased the LPS‐induced cytokines, iNOS, and COX‐2 mRNA expression in RAW264.7 cells, indicating anti‐inflammatory activity. These results indicate that SCLE might be a potential candidate as an anti‐oxidant, anti‐diabetic, and anti‐inflammatory agent.
Transient intrauterine ischemia duringmid‐gestation results in postnatal abnormalities in formation of neocorticalcircuit elements (981.11)
Authors: Sandhu Sonia , Kraushar Matthew , Wijeratne Sagara , Rasin Mladen‐Roko
Published: June 2021

The mammalian neocortex is six‐layered brain structure responsible for higher cognitive functions, voluntary motor behavior, and perception. The differentiation of neocortical projection neurons into each layer requires an uninterrupted coordination of functional gene expression, which may be susceptible to insults like transient intrauterine ischemia (TII). We developed a model of TII in an anesthetized gravid mouse at E15 for 15 minutes. Previously conducted cortical layer‐marker studies of CDP, a transcription factor associated with post‐mitotic dendritic differentiation, showed expanded expression in Ischemic mice relative to Sham mice. Decreased expression of CTIP2, associated with formation of striatal sub‐cortical tracts and L1‐NCAM immunostaining of forebrain axonal pathways in Ischemic brains relative to Sham suggesting further that TII at E15 induces disrupted expression of key genes in the development of neocortical layers. We used Golgi Staining and a digital quantification program to analyze ultrastructure changes in upper and lower layer neurons in both male and female mice in dendritic spine density, dendritic branching, axonal lengths, and cell body size in 8‐week old adult mice for Ischemic and Sham conditions. Remarkably, profound differences in dendritic complexity and dendritic spine density differences were seen postnatally in cytoarchitecture, suggesting that TII is sufficient to cause abnormalities in neocortical circuit elements and implications of neurodevelopmental diseases, such as Schizophrenia.
Lkb1‐ a master tumor suppressor (LB108)
Authors: Aube Eric , Chan Rosanna , Maynard Alex , Ortbals Chloe , Paul Lindsey , Shome Ria , Wright Madison , Kessler Eric , Tuel Kelly , Whalen Joseph , Anant Shrikant
Published: June 2021

The small intestine is remarkable in that 109 epithelial cells are produced every twenty minutes, but cancer rarely develops. This is possible due to the serine/threonine kinase tumor suppressor Liver Kinase B1 (LKB1). Mutations in LKB1 were first linked to the disease Peutz‐Jeghers syndrome in which polyps grow and have a 93% chance of becoming cancerous. Mutations in LKB1 have been linked to several forms of cancer including small intestine, breast, lung, liver, and pancreas. LKB1 forms an active heterotrimer with pseudokinase: STRAD, and scaffolding protein: MO25. LKB1’s primary function is to regulate energy homeostasis through the phosphorylation of the t‐loop structure on adenosine monophosphate protein kinase (AMPK), which mediates the production of ATP when cells enter a state of energy stress. AMPK detects and reacts to differences in the AMP:ATP ratio. AMPK has also been shown to impact several cancer‐related processes. For example, the AMPK‐mTOR signaling pathway is important for inducing cell death within cancer cells. Possible future research includes finding methods of inducing LKB1 activity, which would stimulate the AMPK‐mTOR pathway and diminishes cancer cell proliferation. The CAPS SMART Team (Students Modeling a Research Topic) created physical models of the tumor suppressor protein LKB1 and its target AMPK using 3D printing technology.

Grant Funding Source: Supported by grants from the NIH‐SEPA and NIH‐CTSA.
The effect of food restriction on the expression of some placental proteins (LB24)
Authors: Harrath Abdel Halim , Mansour Lamjed , Al Omar Sulimen , Alwasel Saleh
Published: June 2021

Fast food and ready meals may be an important determinant in the significant increases of numerous diseases, such as reproductive disorders, because these foods are low in macro and micronutrients. In this study, we aimed to investigate whether food restriction in pregnant mothers affects placental structure and function.

The rat represents an excellent organism for studying human disease and the main source for testing new drugs and to increasing knowledge of the pathobiology of human disease. The placentas from pregnant females under food restriction and controls were removed on the day 6, 14 and 20 of pregnancy. A panel of Sulimen Y. Al Omar1differentially expressed proteins, specifically tumor necrosis factor (TNF) signaling, TNF receptor 1 (TNFR1) and transforming growth factor‐β (TGF‐β) have been subjected to immunohistochemistry and molecular analysis. Our observations suggest that the expressions of some of these proteins were affected by the food restriction, and thus placenta is affected by the lack of nutrition during pregnancy. We conclude that these differentially expressed proteins are an effective target for measuring placental defects and related risks to the fetus development. Results from a rat model may be extrapolated to other species including humans.

Grant Funding Source: Supported by Deanship of Scientific Research at king Saud University
Microrna expression distinguish glioblastoma from lower grades astrocytomas (LB286)
Authors: Rivera‐Díaz Mónica , Soto Daniel , Quintero Mario , Marcos María , Vivas‐Mejía Pablo
Published: June 2021

Astrocytomas are the most common type of adult primary central nervous system neoplasms. The World Health Organization (WHO) histologically classifies astrocytomas into four malignancy grades spanning from grade I, a benign tumor, to grade IV, a highly malignant tumor. This classification is highly subjective and more specific molecular markers are needed. MicroRNAs (miRNAs) are small (19‐24 nucleotides) non‐coding RNAs that regulate gene expression at the post‐transcriptional level. Increased in vitro and in vivo evidence indicates that several miRNAs are dysregulated in all cancers types including astrocytomas. However, the precise miRNAs aberrantly expressed in each astrocytoma grade have not been identified. The aim of this study was to elucidate the miRNAs dysregulated in each astrocytoma grade using formalin‐fixed paraffin‐embedded (FFPE) samples. Microarray data was analyzed with the PARTEK software. Eighty‐five miRNAs were differentially regulated in grade II, 72 miRNAs in grade III and 53 miRNAs in GBM compared with control samples. Particularly, miR‐210, miR‐193b and miR‐27a were highly associated with GBMs. Additionally, up‐regulation of miR‐92b and down‐regulation of miR‐1908 occurred in all astrocytoma grades compared with control individuals. Biological pathway analysis showed a relationship between miRNAs signatures and molecular pathways related with tumor initiation and progression of astrocytoma. Taken together, our results provide additional miRNA signatures to distinguish between GBMs and lower grades of astrocytoma.
Effects of Calorie Restriction on Total and Heligmosomoides bakeri‐specific IgG1 in Mice of Differing Immune Phenotypes (LB307)
Authors: Jablanovic Jelena , Lang Samantha , Manibusan Pierre , Kristan Deborah
Published: June 2021

Calorie restriction (CR) is currently being investigated in humans for therapeutic effects on longevity and age‐associated diseases. However, some preliminary animal studies have shown that CR can adversely affect the immune system in the presence of infection; therefore the risk of CR needs to be further evaluated. In this study we subjected 2 immunologically different strains of laboratory mice (C57BL/6, SJL) to CR, and then infected them with intestinal nematode Heligmosomoides bakeri. We then measured a non‐specific immunoglobulin response (total IgG1) and a pathogen‐specific immunoglobulin response (H. bakeri‐specific IgG1). Overall, our study showed that C57 had a more robust total IgG1 response, but H. bakeri‐specific IgG1 was similar in both strains. CR had an attenuating effect on total IgG1 in both C57 and SJL mice, but CR had no effect on the H. bakeri‐specific IgG1 response in either strain. Taken together, our study showed that CR can attenuate a non‐specific immunoglobulin response and have no effect on pathogen‐specific response. However, we caution that effects of CR on pathogen‐specific response should further be investigated during longer or repeated infections.

Grant Funding Source: NIH 5SC3GM084806
Assessing the association between the methylenetetrahydrofolate reductase (MTHFR) 677C→T polymorphism on blood folate concentrations: a systematic review and meta‐analysis of trials and observational studies (LB311)
Authors: Tsang Becky , Cordero Amy , Marchetta Claire , Mulinare Joseph , Mersereau Patricia , Guo Jing , Qi Yan Ping , Berry Robert , Rosenthal Jorge , Crider Krista , Hamner Heather
Published: June 2021

Background: The methylenetetrahydrofolate reductase (MTHFR) 677C→T polymorphism is a risk factor for neural tube birth defects (NTDs). The T allele produces an enzyme with reduced folate processing capacity, which has been shown to produce lower blood folate concentrations in some studies.

Objective: To assess the association between MTHFR C677T genotypes (CC, CT, TT) and blood folate concentrations among women aged 12‐49 years.

Design: We conducted a systematic review of literature published between 1/1992‐7/2013 to identify controlled trials and observational studies that reported serum, plasma, or red blood cell (RBC) folate concentrations and MTHFR C677T genotype. We applied a Bayesian random‐effects model to predict differences in blood folate concentrations between MTHFR C677T genotypes, stratified by folate assay.

Results: Thirty‐eight studies met criteria for inclusion. Serum/plasma folate concentrations showed a consistent genotype trend with the highest concentrations for CC (CC > CT > TT) regardless of assay type. RBC folate concentrations measured by microbiologic assay also demonstrated this trend; however, this trend was reversed (CC < CT < TT) in studies using protein‐binding assays.

Conclusions: Meta‐analyses results showed blood folate concentrations differed by assay type and genotype. Previous evidence has shown that RBC folate concentrations measured with a radioimmunoassay requires adjustment for genotype‐dependent folate recovery; our results suggest that other protein‐binding assays could have similar limitations. Compared to CC individuals, TT individuals have lower blood folate concentrations, which may increase a woman’s risk for an NTD‐affected pregnancy.