bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of student interviews to identify and refine threshold concepts for biochemistry (618.23)
Authors: Lin Sara , Loertscher Jennifer , Minderhout Vicky , Green David , Lewis Jennifer
Published: June 2021

Threshold concepts are fundamental concepts that require mastering in order to progress towards a deeper understanding of a discipline. Despite their importance in teaching and curriculum reform efforts, threshold concepts have not yet been identified for biochemistry. Using an iterative approach involving over 75 faculty members and over 40 students from diverse institutions nationwide, we have developed a list of candidate threshold concepts, which will inform the design of new active learning materials and assessment tools. Student interviews, conducted with biochemistry and organic chemistry students from five different institutions, were essential in refining the pilot list developed by faculty. Each focus group interview probed student understanding of two of the three candidate threshold concepts: dynamic equilibrium, intra‐ and intermolecular interactions, and individual versus populations of molecules. Analysis of interview transcripts revealed wide variability in student understanding of these concepts that could be due to differences in major, prerequisite coursework and/or amount of biochemistry coursework completed. Despite the variability, trends were observed including an inability to articulate the meaning of steady state in a biological context, difficulty articulating the basis of intermolecular interactions in any context, and recognition that moving between thinking about individual versus populations of molecules is an important and challenging part of learning biochemistry.

Grant Funding Source: National Science Foundation NSF‐DUE 1224868
The relative influence of prepregnancy weight and gestational weight gain on offspring birth size in Vietnam (620.7)
Authors: Young Melissa , Nguyen Phuong Hong , Addo O.Yaw , Hao Wei , Nguyen Hieu , Pham Hoa , Truong Truong V , Nguyen Son , Martorell Reynaldo , Ramakrishnan Usha
Published: June 2021

Maternal and child undernutrition remains an important public health problem. The objective of this research is to examine the relative influence of maternal prepregnancy weight (PPW) and gestational weight gain (GWG) on birth size for 971 singleton live births born to women who participated in a pre‐conceptional micronutrient supplementation trial in Vietnam. Adjusted generalized regression models were used to examine associations with birth weight, BW (mean + SD: 3054 + 400 g); small for gestational age, SGA (22%); and low birth weight, LBW (5%). A 1 SD increase in PPW was associated with a 226 g (95%CI: 220‐231) increase in BW. Similar and independent effects were observed for BW (226 g; 95%CI: 219 – 233) per SD increase in GWG. The relative impact of a 1 SD increase in PPW and GWG were similar for length (0.17 and 0.14 cm), head circumference (0.13 and 0.15 cm) and MUAC (0.15 and 0.14 cm) at birth, respectively. A 1 kg increase in PPW and GWG was also associated with a 9.4% and 9.5% reduction in the risk of delivering a SGA infant and 12% and 15% for LBW, respectively. Women with pre‐pregnancy BMI < 17 kg/m2 (moderate‐severe thinness) or in the lower tertile of GWG had 2.4 – 2.6 times the risk for SGA and 3.9 – 4.6 times the risk for LBW. These findings indicate that programs aimed at improving birth outcomes will have the greatest impact if they address maternal nutrition both before and during pregnancy.

Grant Funding Source: Supported by the Micronutrient Initiative & the Mathile Institute for Advancement of Human Nutrition
Comparison of breast milk sodium‐potassium ratio, pro‐inflammatory cytokines, and somatic cell count as potential biomarkers of subclinical mastitis (623.21)
Authors: Wren Hilary , Li Chen , Solomons Noel , Chomat Anne Marie , Scott Marilyn , Koski Kristine
Published: June 2021

Background: SCM is an asymptomatic inflammatory condition of the lactating breast associated with infant growth faltering. A breast milk Na/K ratio >0.6 is the current indicator for SCM whereas in cows SCC is considered the ‘gold standard’. Our objective was to determine if potential biomarkers of human SCM (SCC, pro‐inflammatory cytokines) were associated with Na/K ratio at two stages of lactation.

Methods: Breast milk samples were collected from 53 lactating Mam‐Mayan women with infants < 45d (early lactation) and 52 with infants 4‐6mo (late lactation). Inductively Coupled Plasma Mass Spectrometry measured Na and K. Flow Cytometry was used to measure SCC. Luminex measured 3 pro‐inflammatory cytokines (IL‐6, IL‐8, TNF‐α).

Results: One‐fifth had a breast milk Na/K ratio >0.6; an elevated Na/K ratio was more common in early lactation (26%) compared to later lactation (15%). By contrast, both IL‐8 (10.3 vs 38.3 pg/mL) and SCC (313K vs 457K cells/mL) were lower in early verses later lactation. The Na/K ratio was positively correlated with all 3 pro‐inflammatory cytokines in early lactation whereas only IL‐8 was associated in later lactation. SCC was not correlated with either Na/K ratio or pro‐inflammatory cytokines in either stage of lactation.

Conclusion: Potential biomarkers of SCM vary by stage of lactation. Based on the Na/K ratio, SCM is more prevalent during early lactation. Results suggest that pro‐inflammatory cytokines may emerge as possible biomarkers and that SCC may not be a useful biomarker of SCM in lactating mothers.

Grant Funding Source: McGill University International Mobility Award, NSERC
Perceived relationships between food and acne among a cohort of New York City young adults (635.4)
Authors: Burris Jennifer , Rietkerk William , Woolf Kathleen
Published: June 2021

Anecdotally, many people with acne believe certain foods impact skin health. The objectives of this study were to investigate the perceived relationship between food and acne among young adults in New York City. Participants (n=279, 139 male, 140 female) completed a questionnaire evaluating self‐reported acne severity, food beliefs, and anthropometric and demographic characteristics. One hundred twenty‐six participants (45.2%) reported no acne, 92 (33%) reported mild acne, and 61 (21.8%) reported moderate to severe acne. Among all participants, 59.1% perceived food to influence acne. Chocolate (26.5%), French fries (25.4%) and pizza (24.4%) were the most commonly reported foods believed to aggravate acne. A greater percentage of participants in the acne groups (mild and moderate to severe) reported diet to influence acne severity compared to participants with no acne (p=<0.001). Future research should examine the impact of foods on acne development and severity.
A randomized, double‐blind, placebo‐controlled clinical trial to investigate the efficacy of cranberry fruit powder (Pacran®) in the prevention of recurrent urinary tract infection in women (639.4)
Authors: Fromentin Emilie , Vostalova Jitka , Vidlar Ales , Galandakova Adela , Vrbkova Jana , Ulrichova Jitka , Student Vladimir , Simanek Vilim
Published: June 2021

Cranberry (Vaccinium macrocarpon) is being studied for use in the prevention of recurrent urinary tract infection (rUTI) where it affects the adhesion Escherichia coli to the uroepithelium. This 6 month, randomized, double‐blind, placebo‐controlled trial aimed to evaluate whether a daily dose of 500 mg Pacran® could prevent UTI occurrences in sexually active women with a medical history of at least 2 symptomatic UTI in the previous 12 months. Study participants were randomized to the Pacran® or the placebo group and were followed up for 6 months. The intent‐to‐treat analysis included 176 participants (n=93 in placebo and n=83 in Pacran® group). In the Pacran® group, the proportion of women experiencing at least one UTI was significantly lower than in the placebo group [10.8% versus 25.8%, respectively, p=0.04, with age adjusted 12‐month UTI history (p=0.01), age (p=0.73), and age‐squared (p=0.05) also included in the model]. There was a similar finding when risk of an E. coli UTI was assessed (p=0.03). When the Kaplan‐Meier survival curves were compared, women in the Pacran® group experienced a longer time to first UTI relative to the placebo group (p=0.04). The average number of UTIs per subject was significantly lower in the Pacran® group relative to placebo group (p=0.03). A daily dose of 500 mg Pacran®, proved effective in reducing the risk of symptomatic UTI in women with a history of rUTIs. Pacran® did not influence clinical chemistry, hematology or urinalysis parameters and was well‐tolerated, with no adverse effects reported.
Tomato carotenoid prevention of UVR‐induced damage in human keratinocyte cell lines (645.9)
Authors: Hill Kelly , Canene‐Adams Kirstie
Published: June 2021

Our objective was to determine the potential of tomato carotenoids: lycopene, beta‐carotene, and lutein to protect against ultraviolet radiation (URV) in two normal human keratinocyte cell lines; HaCaT and NHEK. Both skin cell lines were treated with each carotenoid individually, lycopene + beta‐carotene, lycopene + lutein, lutein + beta‐carotene, and all three for a period of 24 hours at dose levels within the ranges seen in human serum. Full‐spectrum UVR (21,700 μW/cm2 UV B‐100 series lamp; Fisher Scientific) was then applied to tissue culture flasks for 20 minutes. HPLC was performed to quantify extent of carotenoid uptake and metabolism. DNA damage was measured using the Comet assay, lipid peroxidation byproducts were measured using the TBARS assay, and inflammation was measured using ELISA kits for TNF‐α, IL‐6, and IL‐10. Results from this study show the potential of carotenoids to protect human keratinocytes against full spectrum UVR damage and cancer‐induction.
Quinoa as source of bioactive compounds with potential for intestinal health (647.18)
Authors: Carrion Rocio , Murphy Kevin , Ganjyal Girish , Kowalski Ryan , Noratto Giuliana
Published: June 2021

Quinoa (Chenopodium quinoa Willd) is a source of nutrients and bioactive compounds with potential to promote intestinal health. Nine quinoa cultivars produced in the State of Washington and one produced in Peru were assessed for their content of non‐digestible bioactive compounds and their stability to cooking using standard analytical methods. We further investigated the prebiotic activity of a cherry vanilla (CV) cultivar with probiotic pills containing Lactobacillus gasseri, Bifidobacterium bifidum and Bifidobacterium longum. Results of extractable phenolics ranged from 1.48 to 2.42 mg/g for the Temuko and CV cultivars and the non‐extractable phenolics ranged from 1.37 to 3.23 mg/g for the Peru and Black cultivars, respectively. A principal component analysis (PCA) showed that the content of bioactive compounds was similar among quinoa cultivars with the exception of Blanca due to its high phytate content. Phenolics and antioxidant activities were more stable in CV and Faro cultivars after cooking, while non‐digestible compounds remained constant only in the CV cultivar. Growth of probiotics was observed in the CV cultivar (1.2‐fold of starch control) and this was accompanied by a decrease in extractable and non‐extractable phenolics by 44.6% and 21.4%, respectively. Overall, these results suggest the potential of quinoa cultivars for intestinal health. We are currently investigating the modulation of gut microflora by CV quinoa consumption using a murine in vivo model for obesity.
Isoflurane induces substrate‐dependent early mitochondrial PTP opening (648.13)
Authors: Agarwal Bhawana , Dash Ranjan , Bosnjak Zeijko , Stowe David , Camara Amadou
Published: June 2021

The volatile anesthetic isoflurane (ISO) may increase permeability of mitochondria to protect against mitochondrial injury. Prolonged opening of the mitochondrial permeability transition pore (mPTP) inevitably causes mitochondrial demise, whereas transient mPTP opening may release accumulated Ca2+ from the matrix to avoid subsequent irreversible mPTP opening due to Ca2+ overload. To test if ISO alters mPTP opening, we isolated mitochondria from Wistar rat hearts to assess buffer Ca2+ uptake by Fura 4F. We found that: (a) ISO (1 mM) induced earlier mPTP opening by increasing Ca2+ uptake rate with succinate (SUC). (b) ISO did not alter mPTP opening but reduced Ca2+ uptake with pyruvate/malate. (c) 10 µM ryanodine or 10 nM imperatoxin A with SUC increased Ca2+ uptake and induced earlier mPTP opening; this suggests that ISO’s effects on mPTP may be due to increased conductance via mitochondrial ryanodine receptors (mRyR). We have patch‐clamped single mRyRs in mitoplasts. Our data infer indirectly that SUC substrate ‐dependent transient mPTP opening may be due to enhanced RyR Ca2+ uptake. Transient Ca2+ release could be a mechanism underlying ISO –induced cardiac protection in ischemia‐reperfusion injury.

Grant Funding Source: P01‐GM066730, R01‐HL095122, R01‐HL089514
Inhibition of β‐catenin transactivation by IFNγ is regulated by Akt1 and 14.3.3z (650.1)
Authors: Candelario Aurora , Nava‐Dominguez Porfirio , Kamekura Ryuta , Laur Oskar , Parkos Charles , Nusrat Asma
Published: June 2021

The proinflammatory cytokine IFNg influences intestinal epithelial cell (IEC) homeostasis in a biphasic manner by acutely stimulating proliferation that is followed by sustained inhibition of proliferation despite continued mucosal injury. B‐catenin activation has been classically associated with increased IEC proliferation. However, we have observed that IFNg inhibits IEC proliferation despite sustained activation of Akt/B‐catenin signaling. Here we show that inhibition of Akt/B‐catenin‐mediated cell proliferation by IFNg requires the formation of a protein complex containing phosphorylated B‐catenin 552 (pB‐cat552) and 14.3.3Z. We identify Akt1 as a bimodal switch that promotes or inhibits B‐catenin transcriptional activity in response to IFNg stimulation. We demonstrate that IFNg initially induces B‐catenin transactivation through Akt‐dependent c‐terminal phosphorylation of B‐catenin to promote its association with 14.3.3Z. Augmented B‐catenin transactivation leads to increased Akt1 protein and active Akt1 accumulates in the nucleus where it phosphorylates 14.3.3Z to induce relocalization of 14.3.3Z/B‐catenin from the nucleus thereby inhibiting B‐catenin transactivation and IEC proliferation. These results outline an unexpected dual function of Akt1 that suppresses IEC proliferation during intestinal inflammation.

Grant Funding Source: Supported by NIH (CP and AN) and Conacyt (PN) grants.
Hypergravity delocalizes thyrotropin receptor (650.3)
Authors: Lazzarini Andrea , Albi Elisabetta , Floridi Antonio , Lazzarini Remo , Loreti Elisabetta , Ferri Ivana , Curcio Francesco , Ambesi‐Impiombato Francesco
Published: June 2021

Thyrotropin receptor (TSHR) is a a heterodimer G protein‐coupled receptor constituted by an α extracellular subunit (53 kDa) interacting with thyrotropin (TSH) and a broad β transmembrane and intracellular subunit (30‐42 kDa) held together by disulphide bridges. TSHR are localized in the basolateral membrane rich in lipid rafts, microdomains with high level of cholesterol and sphingolipids, that provide a platform for the assembly of signaling complexes after TSH‐TSHR interaction. This interaction was perturbed in hypogravity during parabolic flights and longer missions in Space on board the ISS. In the present work the influence of complex mechanical effects of hypergravity on TSHR, cholesterol and sphingolipid in thyroid gland in vivo was investigated. Seven mice were maintained in hypergravity in a 2g centrifuge. Our results show that hypergravity up‐regulated 65% TSHR. TSH treatment did not cause significant changes in the thyroid gland of the control mice and induced about 50% reduction of the α subunit TSHR of 2g animal. TSH‐stimulated cAMP production was similar in control and 2g samples, indicating that even if TSHR is up‐regulated in hypergravity, the loss of its α subunit that binds TSH was responsible for the lack of the expected increased response to hormonal stimulation. Immunofluorescence analysis of TSHR demonstrated that in control samples the receptor was present on the surface of thyrocytes that surrounded follicles with a precise location, whereas in 2g samples the fluorescent signal was higher and spread over the entire surface of thyrocytes. The UFLC‐MS/MS study on lipid fraction of thyroid tissue showed that sphingomyelin and ceramide species remained unchanged whereas cholesterol was reduced 49% in 2g in comparison with control samples. We thus suggest that gravity, by inducing a significant loss of cholesterol, perturbed lipid rafts modifying TSH‐TSHR interaction.

Grant Funding Source: Agenzia Spaziale Italiana
The anti‐oxidant, anti‐inflammatory and antinociceptive activities of the methanol leaf extract of Moringa oleifera in some laboratory animals (657.19)
Authors: Adedapo Adeolu , Falayi Olufunke , Oyagbemi Ademola
Published: June 2021

Moringa oleifera is an annual herbal shrub which has been used in traditional medicine in Nigeria to treat some disease conditions. This study evaluated the methanol leaf extract (MLE) of the plant for pharmacological activities in rats and mice to validate its folkloric use in Nigeria.

MLE was prepared by dissolving ground plant materials (200g) in methanol for 48 h, filtered, dried and thereafter used for pharmacological investigations. Standard phytochemical methods were used to test for the presence of phytoactive compounds in the plant. Anti‐oxidant activities of the plant were assessed using ABTS, DPPH, FRAP and total phenolics test. The anti‐inflammatory activities were conducted using carrageenan and histamine‐induced tests while antinociceptive activities were carried out using writhes and formalin licking tests.

Acute toxicity test showed that no animals died at the doses even at 1600mg/kg dose. The extract at 50, 100 and 200mg/kg body weight caused a significant reduction in the anti‐inflammatory and analgesic activities though not in the same magnitude as indomethacin, the reference drug used in this study. In the case of anti‐oxidant activities, the extract at 20mg caused 86.0% ABTS inhibitory activity; 49.0% DPPH inhibitory activity; 82.7µmol/L ascorbic acid equivalent for FRAP and 17.2µmol/L gallic acid equivalent for total phenolics.

The results from this study may have validated the traditional basis for the use of Moringa oleifera as medicinal agent. The pharmacological activities noted in this study may be attributed to the presence of flavonoids and other phenolics contained in this plant.
Sphingosine 1‐phosphate induces anti‐inflammatory M2 polarization through IL‐4 signaling in macrophages (662.14)
Authors: Park Soo‐Jin , Lee Kyoung‐Pil , Kang Saeromi , Im Dong‐Soon
Published: June 2021

Phenotypes of macrophages, classic pro‐inflammatory M1 and alternative anti‐inflammatory M2, could influence many disease states like atherosclerosis. Lipopolysaccharide (LPS)‐activated macrophages increase pro‐inflammatory cytokine secretion. Such LPS‐induced‐M1 phenotype expression was suppressed by sphingosine 1‐phosphate (S1P), an anti‐atherogenic lipid mediator in HDL. Previously, suppression of NF‐κB activity through S1P1 was suggested as a key mechanism for the suppression of M1 phenotype (Hughes et al. Circ Res. 2008; 102: 950‐958). In the present study, we addressed how S1P induces M2 macrophage polarization in mouse macrophages and found that S1P induced anti‐inflammatory M2 polarization through IL‐4 secretion and its signaling. S1P induced not only induction and secretion of IL‐4 but also induction of their receptors, IL‐4Rα and IL‐2Rγ. In addition, down‐stream signaling such as phosphorylation of stat‐6, induction of SOCS1, and suppression of SOCS3 expression was also observed in response to S1P. Furthermore, S1P‐induced activation of ERK and inhibition of p38 MAPK and JNK were found to be key signaling for IL‐4 induction. Present data suggest that anti‐atherogenic effect of S1P in HDL may be mediated through IL‐4 secretion and its phenotype switching from M1 to M2 anti‐inflammatory macrophages.

Grant Funding Source: This research was supported by Basic Science Research Program through the NRF (2011‐0021158)
N‐cadherin adherens junctions in rat cerebral artery are mechano‐sensitive (664.9)
Authors: Sun zhe , Li Zhaohui , Li Min , Hill Michael , Meininger Gerald
Published: June 2021

N‐cadherins physically link apposing vascular smooth muscle cells (VSMC) via formation of adherens junctions (AJ) that are coupled to the actin cytoskeleton. In isolated rat mid‐cerebral artery, immunofluorescence studies revealed that the density of N‐cadherin‐AJs increased when lumen pressure was raised from 50 to 110 mmHg, suggesting that the N‐cadherin AJs are mechanosensitive and respond to changes of lumen pressure. In addition, the N‐cadherin‐AJs were observed to align along the cell membrane in pattern of strings when pressure increased from 50 to 90 mmHg. Atomic force microscopy (AFM) with N‐cadherin‐coated microbead (10 μm) tipped probes was employed to examine the response of N‐cadherin adhesion sites to pulling force in VSMC. AFM pulling of the N‐cadherin adhesion (~1 nN) induced an opposing force‐generating contractile response from the VSMC. Transfection of freshly isolated VSMC with N‐cadherin‐GFP was used to study clustering of N‐cadherin to the microbead adhesion site. Upon contact with cell membrane, the N‐cadherin‐coated microbead induced progressive clustering of N‐cadherin‐GFP. Furthermore, there was additional recruitment of N‐cadherin to the microbead when AFM pulling force was applied and the bead was pulled back toward the cell surface by the VSMC. Collectively, these results support a functional role of N‐cadherin AJs as mechano‐sensitive sites in the small resistance arteries. (1P01HL095486 to G.A.M.)

Grant Funding Source: Supported by NIH 1P01HL095486 to G.A.M.
Purine metabolism in diving and non‐diving mammals (667.4)
Authors: Lopez‐Cruz Roberto , Barjau Perez‐Milicua Myrna , Crocker Daniel , Gaxiola‐Robles Ramon , Vazquez‐Medina Jose , Zenteno‐Savin Tania
Published: June 2021

When tissues are deprived of blood (ischemia) there is a progressive reduction of oxygen (hypoxia). Tissue function can be maintained until oxygen and ATP stores are depleted. During breath‐hold diving, blood flow to most tissues is decreased and blood is directed preferentially to the central nervous system. Diving mammals (seals and dolphins) routinely dive as part of their natural history. The hypothesis of this study is that diving mammals have a refined purine salvage system to avoid loss of nucleotides and to maximize ATP recycling during ischemia/reperfusion associated to diving. The goal of this study is to analyze the activities of purine nuceoside phosphorylase (PNP), xanthine oxidase (XO) and hypoxanthine‐guanine phosphorybosil transferase (HGPRT), key enzymes involved in the purine salvage pathway in marine mammals adapted to tolerate repeated cycles of ischemia/reperfusion associated with diving (bottlenose dolphin, northern elephant seal, river otter) and compare with those from non‐diving mammals (human, pig). PNP, XO and HGPRT activities were analyzed by spectrophotometry, purine nucleotide metabolite concentrations was analyzed by HPLC. No clear relationship in circulating PNP or XO activity could be established between diving and non‐diving mammals. Intraerythrocyte PNP activity in bottlenose dolphins could be related to a release of purine nucleotides from the liver. The plasma PNP and XO activities observed in river otters may be due to their precarious energetic balance. Low erythrocyte PNP activity and elevated plasma XO activity in northern elephant seal could be due to fasting and/or sleep‐ and dive‐associated apneas.

Grant Funding Source: Supported by CONACYT
Microfluidic device and assay for mimicking the leukocyte‐endothelium interactions in vivo (669.4)
Authors: Lamberti Giuseppina , Prabhakarpandian Balabhaskar , Smith Ashley , Pant Kapil , Wang Bin , Kiani Mohammad
Published: June 2021

There is no in vitro model that allows investigation of the entire leukocyte adhesion cascade. In this study we present a novel microvasculature network based microfluidic device (MFD) that enables a single assay to study the entire cascade.

Our modified Geographic Information System (GIS) approach was used to digitize the microvascular networks and generate the MFD using soft‐lithography based microfabrication. The MFD consisted of vascular channels seeded with endothelial cells, in communication with the tissue compartment filled with chemoattractant (fMLP) via a 3µm porous barrier. Leukocytes were flowed on activated endothelial cells under physiologic flow conditions.

Leukocytes rolling, adhesion and migration was imaged as a function of time and position in the MFD and compared with in vivo data from mouse cremaster muscle. Our findings indicate that in MFD rolling and adhesion patterns are in agreement with the in vivo data, with 50% of adhered leukocytes migrating from the vascular channels into the tissue compartment after 120min. Furthermore, we blocked the individual steps treating endothelial cells with E‐selectin and ICAM‐1antibody and leukocytes with wortmanin, showing that we can respectively reduce rolling to ~20%, adhesion to ~50%, and migration to ~18% of their original values. In conclusion, this device more realistically mimics geometric and physiologic characteristics of an in vivo microvascular network for studying the entire leukocyte cascade during inflammation.

Grant Funding Source: Supported by NIH
Upregulation of pro‐inflammatory cytokine receptors in forebrain subfornical organ and hypothalamic paraventricular nucleus in rats with heart failure (686.23)
Authors: Wei Shun‐Guang , Zhang Zhi‐Hua , Yu Yang , Weiss Robert , Felder Robert
Published: June 2021

The pro‐inflammatory cytokines tumor necrosis factor‐α (TNF‐α) and interleukin (IL)‐1β play an important role in regulating sympathetic drive and neuroendocrine activation in heart failure (HF). Previous studies in normal rats have demonstrated that receptors for TNF‐α and IL‐1β are abundant in the subfornical organ (SFO) but only sparsely expressed in the hypothalamic paraventricular nucleus (PVN). In HF rats, TNF‐α and IL‐1β increase in the circulation and in cardiovascular autonomic regions of the brain. Pro‐inflammatory cytokines can upregulate their own receptors, but the question of whether TNF‐α and IL‐1β receptors increase in SFO or PVN in HF has not been addressed. We examined TNF‐α and IL‐1β receptor (IL‐1RAcP component) expression in SFO and PVN in adult male Srague‐Dawley rats 4‐5 weeks after myocardial infarction or sham operation (Sham), using immunofluorescent staining and real‐time PCR analysis. Fluorescent intensity for TNF‐α and IL‐1β receptors was significantly (*p<0.05, n=6) greater in both SFO and PVN in the HF rats, compared with Sham rats. Similarly, the mRNA levels of TNF‐α and IL‐1β receptors in SFO and PVN were significantly (n=5) higher in HF compared with Sham rats. These data indicate that upregulation of TNF‐α and IL‐1β receptors contributes to the inflammatory state driving sympathetic and neuroendocrine activity in these key cardiovascular autonomic regions of the brain in HF.

Grant Funding Source: Supportd by NIH R01HL073986
Adipose dysfunction with aging is associated with resistance artery endothelial dysfunction and impaired angiogenic capacity (688.3)
Authors: Hazra Sugata , Henson Grant , Bramwell R , Reihl Kelly , Donato Anthony , Lesniewski Lisa
Published: June 2021

Although tissue dysfunction is reported in aging, effects of aging on adipose tissue per se are not well studied. To address this, we assessed metabolic homeostasis, adipose tissue structure‐function and adipose artery endothelial function from young (6 mo) and old (29 mo) male B6D2F1 mice. Fasted blood glucose was not different with age but plasma insulin, homeostatic model assessment (HOMA‐) of insulin resistance (IR), and beta cell function (‐B) were increased by 84%, 54%, 57% respectively (P<0.05) in old mice. Old mice also were glucose intolerant (P<0.05), concomitant with increased plasma free fatty acids (P<0.05) and liver triglycerides (P<0.05). Despite no difference in body mass, epididymal white adipose tissue (eWAT) mass was 40% lower in old mice. There was ~60% reduction in mean adipocyte area (p<0.01), 7‐fold increase in fibrosis (p<0.01) and ~50% reduction in vascular density (p<0.01) in old eWAT. There also was a 2‐fold reduction in in vitro eWAT angiogenesis and a ~25% reduction in endothelium dependent dilation to acetylcholine in eWAT arteries (p<0.01) from old mice, without a reduction in endothelium independent dilation. Thus, aging results in metabolic dysfunction associated with increased circulating and ectopic lipids, altered adipose tissue morphology and reduced vascular density, concomitant with impaired angiogenic potential and endothelial dysfunction in adipose arteries.

Grant Funding Source: Supported by R21 AG033755, K01 AG033196, R01 AG040297
The effects of streptozocin‐induced type I diabetes on P2 receptors profile in the podocytes of the Dahl SS rat glomeruli (689.6)
Authors: Lowing Andrea , Ilatovskaya Daria , Palygin Oleg , Staruschenko Alexander
Published: June 2021

Calcium flux in the podocytes is critical for their physiological function; excessive calcium in these cells can result in glomeruli damage, proteinuria and reduced GFR. P2 receptors are located throughout the kidney, and upon binding of ATP they activate calcium influx that triggers a plethora of intracellular processes. Our previous data have demonstrated that the major P2 receptor in the Sprague Dawley rat podocytes is P2Y1. Diabetic conditions have been shown to cause glomeruli injury and alter purinergic receptors profile in the kidney. The goal of the current study was to identify the P2 receptors responsible for calcium flux in the podocytes of the rats with type I diabetes. Type I diabetes was induced in the Dahl salt‐sensitive (SS) rats by an injection of streptozocin followed by an insulin implant, which results in the development of hyperfiltration, progressive proteinuria and renal damage typical for diabetic nephropathy. After 6 weeks of treatment, the glomeruli of the rats were isolated by differential sieving, loaded with Fluo‐4/FuraRed calcium dyes and analyzed with real‐time ratiometric fluorescent measurements. Pharmacological agonists and antagonists of the P2X and P2Y receptors were utilized to determine P2 receptors profile under diabetic conditions. This study will define the P2 receptors involved in the pathogenesis of the glomerular injury in diabetes, which could lead to a targeted treatment for renal complications associated with this disease.
Beneficial effect of Oryeongsan in diabetic nephrosclerosis (701.2)
Authors: Yoon Jung Joo , Lee Yun Jung , Lee So Min , Kim Mi Kyeong , Kang Dae Gill , Lee Ho Sub
Published: June 2021

Beneficial effect of Oryeongsan in diabetic nephrosclerosis

Jung Joo Yoon1,2, Yun Jung Lee1,2, So Min Lee 1,2, Mi Kyeong Kim1,2, Dae Gill Kang1,2*, Ho Sub Lee1,2*

1College of Oriental Medicine and Professional Graduate School of Oriental Medicine, 2Hanbang Body‐fluid Research Center, Wonkwang University, Shinyong‐dong, Iksan, Jeonbuk, 570‐749, Korea

Mesangial fibrosis and expansion are caused by extracellular matrix accumulation leading to diabetic nephrosclerosis. Oryeongsan, traditional herbal formula, is widely used for the treatment of nephrosis, dropsy, and uremia. In this study, the anti‐fibrotic effect of Oryeongsan was investigated in db/db mice.

Db/db mice were treated with 100 mg/kg Oryeongsan daily for 8 weeks. In Oryeongsan treated db/db mice, plasma glucose/insulin, glucose tolerance, and HOMA‐IR index were significantly improved compared with untreated db/db mice, respectively. Immunohistological analysis revealed that glomerular type IV collagen and TGF‐β1 expression was decreased by Oryeongsan. Oryeongsan was also markedly reduced the ratio of mesangial matrix/glomerular area. In addition, protein expression levels of TGF‐β1, Smad‐2/‐4, type IV collagen, CTGF and TIMP were significantly decreased by Oryeongsan, whereas, Smad‐7 and MT1‐MMP were increased in the renal cortex. Smad‐2/‐4 activation in nucleus was also markedly decreased by Oryeongsan.

Therefore, this study suggests that Oryeongsan suppresses mesangial fibrosis through the inhibition of type IV collagen and TGF‐β1/Smads action. Hence, Oryeongsan may be a new therapeutic option for the treatment of diabetic nephrosclerosis.
The effect of light load resistance exercise and whey protein supplementation on muscle protein synthesis and amino‐acid transporters in elderly (702.7)
Authors: Agergaard Jakob , Bülow Jacob , Kildevang Jensen Jacob , Reitelseder Søren , Schjerling Peter , Drummond Micah J. , Kjær Michael , Holm Lars
Published: June 2021

In this study we tested how light load resistance exercise (LL RE) affects skeletal muscle protein synthesis (FSR) and amino‐acid transporter (AAT) protein expression as a way to counteract anabolic resistance and age related loss of muscle mass. Untrained healthy men (age: +65 yrs) were subjected to 13 hours supine rest. After 2½ hours of rest, unilateral LL RE was conducted: 10 times 36 reps of knee‐extensions at 15% 1RM. Hereafter, the subjects were randomized to oral intake of Placebo (4g maltodextrin/hour) (n=10), Pro‐C (4g whey protein/hour) (n=10) or Pro‐2 (28g whey protein at 0 hours and 12g whey protein at 7 hours post exercise) (n=10). Quadriceps muscle biopsies were taken at 0, 3, 7 and 10 hours post exercise from both resting and exercised leg. Myofibrillar‐FSR and membrane protein expression of select AATs were analyzed from the biopsies. LL RE increased myofibrillar‐FSR compared to the resting leg in all groups. An increase in AAT protein expression was only observed when LL RE was followed by whey protein intake. Specifically, Pro‐C increased LAT1, PAT1 and a tendency towards increased SNAT2 protein expression, Pro‐2 only increased PAT1 protein expression. We conclude that myofibrillar‐FSR increased in response to LL RE, irrespective of feeding in older adults. AAT protein expression only increased when LL RE was combined with whey protein intake.

Grant Funding Source: Arla Foods Ingredients Group P/S, Viby J, Denmark
Electronic whiteboard‐style instructional videos on the cheap (720.4)
Authors: Pressley Thomas , Fowler John
Published: June 2021

Instructional videos are an increasingly popular format for delivering content to students. Although there are a variety of medical subjects available, we found that few existing videos were relevant or appropriate for teaching first‐year medical students. Moreover, we wanted our own institution to be well represented in any collection of videos that we recommended to our class. Among various presentation strategies that we explored, the electronic whiteboard‐style video popularized by the Khan Academy is a good match for material that our faculty might traditionally present by freehand drawing on the blackboard, such as simple diagrams or the derivation of equations. We found that we could develop such videos by taking advantage of relatively‐inexpensive hardware and software. Each video was planned using traditional storyboarding techniques and storyboard templates available for free online. We then recorded the narration digitally using inexpensive headsets and free software such as Audacity. With the recorded narration and storyboard as a guide, video of the drawings was recorded using digital tablets, drawing software such as Paint or SmoothDraw, and screen‐capture utilities such as Expression Encoder or Screenr. The hardware and software were inexpensive, often already in use by faculty, or were freely‐available for download. Finally, modestly‐priced video editing programs such as Camtasia Studio or CyberLink PowerDirector were used to join the captured drawings and audio into a single video, which could then be posted to a learning management system for streaming to students. Our assessment strategy consists of comparisons of examination performance on concepts covered in these videos with previous years that covered the same material in traditional lecture or small group. These comparisons are supplemented with pre‐ and post‐instruction surveys of student opinion on the effectiveness of the videos in improving their studies and interviews with participating faculty.
Developing a comprehensive graduate‐level knowledge base through a functional histology course experience (721.17)
Authors: Swailes Nathan , Heidger Paul
Published: June 2021

At Iowa, a comprehensive graduate‐level course in General Histology is offered for basic science graduate students in the face of diminishing time allocations for Histology in the medical curriculum. It consists of 24 hours of general lecture and 37 hours of supervised virtual and optical microscope laboratory experiences, offered in tandem with the required histology component of our medical Structure and Function of Human Organ Systems course. In addition, graduate students are mentored weekly in student and faculty‐led discussion sessions emphasizing advanced topics, the current research literature, microscopic technique, and histopathologic relevance to each student’s research area. Course requirements are rigorous, and include active, creative participation in the advanced topics sessions and achieving at least 80% on all lecture and lab examinations. Didactic exams include both essay questions and image/non‐image linked multiple choice questions that are designed to assess an integrated knowledge of structure and function relationships. Mastery of laboratory objectives is assessed each unit using virtual image‐based practicals which emphasize cell and tissue detail; an oral lab exercise at the microscope with an instructor, emphasizing structure/functional relationships; and a comprehensive final glass slide exam emphasizing organ identification. Collectively, these assessments ensure students have achieved competency in microscope skills and a functional understanding of cells, tissues, and organs. Student feedback confirms that their learning experience prepares them well to fulfill with distinction their teaching assistant requirement for graduation, and to assume full responsibility in their future faculty teaching roles.
Virtual histology laboratory with an interactive tutorial feature: transitioning into the future of histology education (725.11)
Authors: Howery Hollis , Lee Lisa
Published: June 2021

The objective of the project was to create a beta version of a virtual histology laboratory website incorporating an interactive tutorial feature that can effectively simulate an in‐class interaction with an expert. Although many virtual histology laboratories already exist and have shown their effectiveness in reducing cost and space while increasing instructional efficiency, few can be classified as a successful stand‐alone learning tool. Learning histology requires pattern recognition skills acquired through hands‐on, instructional sessions conducted by an expert identifying key structures, quizzing students and providing feedback. The interactive tutorial feature simulates this expert interaction through an overlay of annotated identification markers on characteristic histological structures and self‐assessment tool which provides instant feedback at a click of a button. This innovative add‐on feature to the virtual histology laboratory is expected to help students obtain mastery in histological pattern‐recognition on their own, and be less dependent of an expert’s presence. A beta version of the website will be available for exploration of its features and effectiveness as a stand‐alone learning tool.
Investigating oxidative modification of anti‐oxidant enzymes and protein carbonylation as mechanisms of cigarette smoke‐induced cardiac stem cell damage (732.6)
Authors: Sumanasekera Wasana , Dao Halle , Gyamfi Felix , Shultz Kadi , Le Uyen , Rokosh Gregg
Published: June 2021

Rationale and Objective: Cigarette smoking is a major health crisis across the globe resulting in many diseases including cardiovascular injuries. This study mainly focused on delineating mechanisms of CS induced cardiac stem cell (CSC) damage, which was observed earlier by our research group. We hypothesized that cigarette smoke induced reactive oxygen intermediates modify anti oxidant enzymes and proteins present in CSC via carbonylation leading to CSC damage, which can be attenuated by ascorbate treatment. Methods: C‐kit positive CSCs have been used as the experimental model. Following treatment of CSCs with cigarette smoke extract (CSE) for 24 hours, CSE induced oxidative modification of CSC proteins, total glutathione level, catalase activity, inhibition of super oxide dismutase (SOD) activity and ascorbate mediated prevention were assessed. The employed methods were; oxi‐ELISA, oxi‐blot / SDS polyacrylamide electrophoresis followed by western blotting, catalase, SOD activity, and total glutathione detection assays. Results: CSE induced oxidative modification / carbonylation of CSC proteins, inhibition of SOD activity, and alleviation of inhibition by ascorbate was observed. CSE caused increase in both catalase activity and total glutathione level in CSCs, which are indicative of oxidative stress. Ascorbate treatment decreased catalase activity indicating squelching of free radicals. Ascorbate increased total glutathione level in CSCs. Conclusions: Oxidative stress induced modulation of CSC proteins and antioxidant enzyme activity is the main mechanism for CSE induced attenuation of CSC function. Ascorbate served as a preventative measure. Implications: As CS attenuates CSC functions, it is crucial to find a way to optimize CSCs prior to transplant. Ascorbate pre‐conditioning serves as a promising target.

Grant Funding Source: Supportyed by Sullivan University Faculty Development Grant
Comparison of lipoplasty technique effects on the cellular fate potential of adipose derived stem cells (732.7)
Authors: Belogorodsky Dimitry , Genoese Angela , Debo Matthew , Murphy Daniel , Chmielewski Sarah , Cho Ellen , Adams Christopher , D'Angelo Marina
Published: June 2021

Clinical use of stem cells provide undeniable advantages as they can substitute for injured cells, tissues, and even organs in the human body. Adult adipose‐derived stem cells (ADSCs), have vast potential for clinical medicine because they can easily be harvested from a patient and provide an autologous transplant option. Data gathered in our lab has shown that harvesting ADSCs using water‐jet assisted liposuction (BodyJet) with laser‐assisted lipoplasty (SmartLipo) techniques produces a high yield of viable stem cells. These isolated cells can be plated in monolayer culture and retain their stem cell phenotype. In this study, we test the differentiation abilities of these BodyJet/SmartLipo harvested cells following cryopreservation. Since the clinical use of ADSCs is highly indicated for orthopedic and arthritis applications, we tested the ability of the liposuction‐harvested ADSCs to differentiate along the chondrocyte and osteoblast lineages. In this study, we compare the cryopreserved primary human ADSCs to a normal human, mesenchymal ADSC line, to measure stem cell viability and differentiation following thaw. Utilizing confocal microscopy, real‐time PCR, cell counts, alizarin red and alcian blue staining, the stem cells show promise in their ability to differentiate along the chondrocyte and osteoblast lineages. These data support the clinically‐relevant idea that lipoplasty‐obtained ADSCs will retain multipotency.

Grant Funding Source: Supported by Pennsylvania Department of Health
Cellular and molecular characteristics of neonatal skin: consequences for healing (734.10)
Authors: Krejci Eliska , Dvorankova Barbora , Kvasilova Alena , Borsky Jiri , Grim Milos , Smetana Karel
Published: June 2021

Wound healing is a complex and dynamic process of restoring tissue integrity and layers of skin. While in adults healing results in replacement of normal skin structures with scar tissue, the process is scarless during initial periods of prenatal development. Reduced scar formation was also observed shortly after birth.

Having the opportunity to obtain newborn skin after corrective surgery of cleft lip, we aimed to compare properties of cellular elements from neonatal and older skin that could be responsible for nearly scarless healing.

Cultivated neonatal fibroblasts expressed Nestin, a marker of immature cells, and markers of stem cells Nanog, OCT3/4, SOX2. Neonatal fibroblasts also retained some degree of plasticity as they could be differentiated into adipocytes, smooth muscle cells and chondroblasts but not to osteoblasts.

Cultivated neonatal keratinocytes included population of very small cells (4 µm in diameter) exhibiting both epithelial (keratin) and mesenchymal (vimentin) marker, as well as some epidermal stem cell markers.

These preliminary data suggest that minimal scarring of neonatal skin could be at least partially explained by persistence of some prenatal properties of cells that retained stem cell characteristics.

Grant Funding Source: Supported by GACR project nr. 13‐20293S, PRVOUK project nr. P27/LF1/1 and UNCE project nr. 204013.
Mechanism of mismatch‐provoked excision in a defined mismatch repair system (735.14)
Authors: Lee Sanghee , Gu Liya , Yang Wei , Li Guo‐Min
Published: June 2021

DNA mismatch repair (MMR) is a highly conserved genome‐maintenance pathway that ensures replication fidelity by targeting repair of replication‐induced mispairs to the newly‐synthesized strand. Despite that both the bacterial and the human MMR reactions have been reconstituted in vitro using purified proteins, many fundamental questions in MMR remain unanswered, one of which concerns the mechanism by which MMR proteins facilitate communication between the mismatch and a distant strand discrimination signal (a single strand nick). At least two models have been proposed to address this issue. The moving model suggests that mismatch recognition protein (MutS in E. coli and MutSα or MutSβ in eukaryotes) slides along the DNA helix from the mismatch to the strand break, where an exonuclease is recruited to initiate mismatch removal. However, the stationary model emphasizes that MutS proteins, while staying bound at the mismatch, interacts with other MMR proteins loaded at the strand break to induce DNA bending or looping that brings the two distant sites together. This long‐standing controversial issue remains unresolved. Here, we tested these models in vitro, using a defined MMR system and a circular plasmid DNA substrate containing a mismatch and physical 'roadblocks' that inhibit MutS protein sliding. We demonstrate that mismatch‐provoked excision proceeds efficiently from the nick even in the presence of the roadblocks, indicating that initiation of excision is independent of whether MutS proteins slide from the mismatch to the nick. This result implies that the communication between the mismatch and the nick is likely through interactions between the mismatch‐bound MutS proteins and other MMR components at the strand break. Our data therefore support the stationary model.