bluuebunny/crossref_metadata_2025 huggingface dataset:
Loss of endogenous H2O2‐induced inhibition of ASIC1‐mediated Ca2+ influx in pulmonary artery smooth muscle cells following chronic hypoxia
Authors: Plomaritas Danielle R , Herbert Lindsay M , Resta Thomas C , Walker Benjimen R , Jernigan Nikki L
Published: June 2021

Our laboratory has demonstrated an important role for acid‐sensing ion channel 1 (ASIC1) in mediating increased pulmonary arterial smooth muscle cell (PASMC) Ca2+ influx and enhanced agonist‐induced vasoconstriction following chronic hypoxia (CH)‐induced pulmonary hypertension. ASIC1 activity is highly sensitive to changes in the redox state of the cell; in particular, hydrogen peroxide (H2O2) has been shown to inhibit ASIC1 function. Furthermore, alterations in reactive oxygen species are known to contribute to the pathogenesis of pulmonary hypertension. Therefore, we hypothesize the increased ASIC1‐mediated store‐operated calcium entry (SOCE) in PASMCs following CH is due to decreased H2O2, attributable to a down regulation of superoxide dismutase (SOD). We found that PEG‐catalase augmented ASIC1‐dependent SOCE in PASMCs from control rats, but was without effect on SOCE in PASMCs from CH rats (4 wks @ 380 Torr). However, the addition of H2O2 inhibited SOCE in both groups. Additionally, SOD1 and SOD3 protein expression were decreased in isolated pulmonary arteries from CH rats compared to control. Together these data suggest that downregulation of SOD1 and SOD3 following CH results in a loss of endogenous H2O2‐induced inhibition of ASIC1‐dependent SOCE in PASMC. This work is supported by NIH HL‐09258 (NLJ) and HL‐07736 (BRW).
Alterations in Intracellular Ca2+ Release via CD38‐Dependent Pathways in Pulmonary Arterial Smooth Muscle of Rat by Chronic Hypoxia
Authors: Lee Suengwon , Jiang Yongliang , Paudel Omkar , S James , Sham K.
Published: June 2021

CD38 is a multifunctional enzyme for synthesis of cyclic ADP‐ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP), which activate ryanodine receptors of sarcoplasmic reticulum and NAADP‐sensitive two‐pore channels (TPCs) of lysosomal stores, respectively, leading to intracellular Ca2+ release. It is thought to play important roles in regulation of intracellular Ca2+ concentration ([Ca2+]i) and vascular functions. Ca2+ homeostasis in pulmonary arteries (PAs) is altered during the development of pulmonary hypertension by chronic hypoxia (CH). However, it still remains unclear whether those CD38–dependent pathways are affected. Here, we examined the changes in the expression of CD38 and TPCs in endothelium denuded PA, and angiotensin II (AngII)‐induced Ca2+ release (AICR) in PA smooth muscle cells (PASMCs). The expressions of CD38 and TPCs in PAs of CH rats were significantly increased. In addition, AICR was found to be mediated in part via the CD38‐dependent pathways. AICR in PASMCs with hypoxic exposure was significantly enhanced compared to control and was reduced by various types of inhibitors for the CD38‐dependent pathway. In summary, our results show that CH exposure leads to changes in expression of CD38 and TPC1/2 in PA smooth muscle as well as increased their associated responses in PASMCs, which may contribute to the alterations in Ca2+ homeostasis. (Supported by R01‐HL075134)
Stable Expression of the Sodium Iodide Symporter (NIS) in Metastatic Cancer Cells: A Novel Imaging Tool
Authors: Gagnon Kenneth Bradley , Chapman Dean , Cooper David M. L. , Caine Sally , Verge Valerie , Nichol Helen
Published: June 2021

In most cancer patients, tumor metastasis to distant organs is the leading cause of death. Tumor cells dissociated from the primary site generally first encounter the lung, making it a significant site of tumor metastasis. We have generated a construct containing the cDNA for the rat sodium iodide symporter (NIS) and stably expressed it in mouse melanoma (B16F0) cells. Intravenous injection of these genetically‐modified B16F0 cells resulted in black tumor nodules throughout the lungs after 14–21 days. Hematoxylin and eosin staining showed numerous mitotic figures in these tumor nodules indicating cell proliferation. In addition, tumor cells were found surrounding capillaries suggesting both angiogenesis and possible routes for metastasis. High resolution in vivo imaging of cancer tumors in large opaque animals is extremely difficult. The high energy X‐rays produced by synchrotron light are capable of overcoming this obstacle and producing in vivo images of these metastatic tumors with much greater sensitivity. Thus, in this study we have developed a novel animal model using these modified tumor cells to accumulate iodine as a contrast agent for in vivo synchrotron imaging of metastatic lung cancer.
Prostaglandin E2 EP3 receptor modulation of insulin secretion in diabetes
Authors: Ceddia Ryan P , Gannon Maureen , Breyer Richard M.
Published: June 2021

Prostaglandin E2 (PGE2) impairs glucose stimulated insulin secretion (GSIS) through its cognate G‐protein coupled receptors (GPCRs). Inhibition of EP3, the predominant PGE2 GPCR in the pancreas, was investigated as a means of improving islet function and diabetic phenotype. The role of PGE2 and EP3 in GSIS was assessed ex vivo with static islet culture utilizing EP3 antagonist, DG‐041, and islets from EP3−/− mice. EP3 blockade improved GSIS in the presence of PGE2. Wild type (WT) and EP3−/− animals on control feed had similar body weights and glucose tolerance. A high‐fat diet (HFD)‐fed model of type 2 diabetes was used to investigate the effects of EP3 inhibition in vivo. Acute DG‐041 administration to WT mice on HFD improved glucose clearance. When place on a HFD, EP3−/− animals gained more weight and developed more severe insulin resistance than did WT mice. EP3−/− mice on HFD had impaired glucose handling compared to WT. These results suggest that acute EP3 blockade improves glycemic control by increasing GSIS, while chronic global EP3 deletion increased obesity and insulin resistance. Funding for research was provided by a grant from the NIH, DK37097.
Opioid‐mediated sustained ligand‐activated preconditioning induces cardiac protection in type II diabetes mellitus
Authors: Hoe Louise Elizabeth See , Zemljic‐Harpf Alice , Schilling Jan , Finley Cameron , Kellerhals Sarah , Roth David , Peart Jason , Patel Hemal , Headrick John
Published: June 2021

Classic preconditioning is impaired in aged and diabetic hearts; however, opioid‐mediated sustained ligand‐activated preconditioning (SLP) is effective in aged myocardium. Here we test cardioprotective effects of SLP in a mouse model of type II diabetes (T2D) induced by streptozotocin with 2 months of high fat feeding. T2D mice exhibited markedly elevated body weight (30%) and fasting glucose, decreased ability to handle glucose loading, and myocardial dysfunction on echocardiography.

Analysis of isolated perfused hearts revealed substantially greater contractile dysfunction and LDH efflux, following ischemia‐reperfusion in T2D vs. non‐diabetic hearts. SLP (5 days subcutaneous morphine pellets) reversed T2D contractile dysfunction in vivo and enhanced ex vivo I‐R tolerance to levels observed in non‐diabetic hearts (also protected by SLP), in association with significant improvements in Ca2+‐dependent mitochondrial swelling and respiratory function, with suppressed ROS generation. SLP also normalized blood glucose and induced a 20% fall in body weight.

In summary, T2D‐dependent cardiac dysfunction and ischemic intolerance are reversed by SLP in association with mitochondrial preservation and shifts in glucose homeostasis and body weight. SLP may be a novel intervention in disease states refractory to conventional approaches.

Research support provided by NIH, VA Merit and QLD Intl Fellowship.
Short‐term estrogen depletion does not promote hypertension in mice
Authors: Fleming J. Daniel , Gray William R. , Liu Shi J. , Hill Brent J.F.
Published: June 2021

Our lab has previously demonstrated that estrogen (E2) downregulates the expression and function of voltage‐gated, L‐type Ca2+ channels (CaL) in coronary arteries in vitro. In this study we hypothesized that the in vivo depletion of E2 would generate an increase in Ca2+‐dependent arterial tone. Mice (C57BL/6) underwent an ovariectomy (OVX) or sham surgery at 8‐wks of age. Biotelemetry measured a drop in heart rate (p=0.002) and blood pressure (p=0.09) in OVX mice within 2‐wks post‐surgery suggesting a baroreceptor reflex compensation. Mice were sacrificed 4‐wks post‐surgery and the mesenteric arteries isolated. The OVX mice demonstrated atrophied uteri and low E2 plasma levels. In the mesenteric arteries, Western blots demonstrated a slight (p=0.12) increase in CaL expression from OVX mice. Mesenteric arterial reactivity experiments indicated no difference between groups in their spontaneous tone and response to the CaL agonist, FPL64176 (3×10−7 to 1×10−6). This suggests that, while E2 depletion alone does not promote hypertension, other underlying pathologies in conjunction with E2 depletion may lead to hypertension and other cardiovascular diseases. Support: NIGMM of the NIH, Grant #P20 GM103429–11
Cardiac Dysfunction in the 5/6 Ablation‐Infarction Model of Chronic Kidney Disease
Authors: Kuczmarski James M , Martens Christopher R , Lennon‐Edwards Shannon L , Edwards David G
Published: June 2021

The purpose of this investigation was to study cardiac function in a rodent model of chronic kidney disease (CKD). Cardiac function was measured in 5/6 ablation‐infarction (5/6 AI) and sham male Sprague Dawley rats 8‐weeks post‐surgery using an isolated working heart preparation. In addition, heart mass was determined and left ventricular (LV) tissue was used for biochemical analysis. Cardiac output was impaired in 5/6 AI at left atrial filling pressures of 13.5cmH2O (50±3 vs. 28±5ml/min; mean ± SEM), 17.5cmH2O (53±6 vs. 29±5ml/min), & 21.5cmH2O (53±6 vs. 30±5ml/min; all p<0.05), but not 9.5cmH2O (39±3 vs. 24±5ml/min; p>;0.05), while aortic pressure was maintained at 80cmH2O. Cardiac output was also impaired at aortic pressures of 60cmH2O (43±3 vs. 24±4ml/min) & 70cmH2O (45±3 vs. 24±4ml/min) with filling pressure set at 13.5cmH2O (all p<0.05). Heart mass was increased in 5/6 AI (3.5±0.3 vs. 4.9±0.4mg/g) and accompanied by increased LV NOX‐4 (1.0±0.1 vs. 1.5±0.1 relative to β‐actin p<0.05) and H2O2 (179±15 vs. 230±17nm/mg). The LV Nox‐2 subunit gp91phoxwas not increased in 5/6 AI (1.0±0.1 vs. 1.1±0.1 relative to β‐actin p>;0.05). In conclusion, cardiac hypertrophy & dysfunction are evident in CKD. These findings could be, in part, mediated by increased Nox‐4 and reactive oxygen species (ROS) production. However, further investigation is needed to clarify the role of NOX‐4 and ROS in uremic cardiomyopathy.
Metabolic and Coronary Dysfunction in Obesity Resistant and Obesity Prone High Fat Fed Rats
Authors: Oltman Christine L , Dake Brian L.
Published: June 2021

The purpose of this study was to compare physiologic, metabolic and microvascular complications in obesity resistant (OR) and obesity prone (OP) rats fed a high fat (HF) diet. 8 week old male rats were fed a diet containing 45% kcal saturated fat for 42 weeks. Blood pressure was measured by telemetry. Reactivity of small (75 – 150 μm intraluminal diameter) coronary arteries was evaluated. Body weights were 504±7, 669±25*, 685±8*, and 924±39* grams for OR control, OR HF, OP control OP HF rats, respectively. Blood pressure was increased in OP rats on control diet (121.5 ± 4.3mmHg*) and further increased with HF diet (129.7 ± 2.4mmHg*), compared to OR rats on control (98.3 ± 4.3mmHg) or HF diet (99.5 ± 3.1mmHg). Glucose clearance was impaired in high fat fed OR and OP rats. Acetylcholine (ACh)‐mediated relaxation of coronary arteries from HF fed rats was impaired (% relaxation to 10 μM Ach: OR control=67 ± 10, OR HF = 36±12*, OP control = 41±8*, and OP HF = 22±6*). Sodium nitroprusside (SNP) responses were not altered (% relaxation to 0.1 mM SNP: OR control = 72 ± 5, OR HF = 60 ± 7, OP control = 62 ± 8, and OP HF = 74 ± 6%). *p<0.05 vs OR control diet. These studies indicate that HF diet augments metabolic and microvascular complications in OP rats.

Supported by VA Merit to Dr. Oltman.
Skin and cerebral vascular function/reactivity in healthy young adult african americans
Authors: Kim Kiyoung , Hurr Chansol , Harrison Michelle L. , Lee Joshua F. , Christmas Kevin M. , Brothers R. Matthew
Published: June 2021

African Americans (AA) are at an increased risk for cardio and cerebral vascular disease relative to Caucasians (CA) and the underlying impairments manifest as early as the second generation prior to overt signs of risk. The mechanisms of this increased risk are multifactorial; however, evidence suggests that microvascular dysfunction is a primary contributor. This study tested the hypothesis that microvascular function, indexed by the skin vascular conductance (SkVc) response to local heating, is impaired in young otherwise healthy AAs. Furthermore, we hypothesized that AAs have an attenuated cerebral vasodilator response to hypercapnia. To date complete data sets have has been collected on 5 AAs and 6 CAs. SkVc was assessed while the skin was clamped at 34 °C and 40 °C and values were normalized to a maximal value obtained during heating at 43 °C for 30 min. Cerebral vasomotor reactivity (CVMR) was assessed by increases in cerebral vascular conductance (CVC) during a rebreathing protocol. CVC was reduced in the AAs at 34 °C (AA: 10±3 % max vs. CA: 20±8 % max; P=0.01) while there was a trend for CVC to be reduced at 40 °C (AA: 57±19 % max vs. CA: 71±8 % max; P=0.12). CVMR was significantly reduced during hypercapnic rebreathing in the AAs relative to the CAs (AA: 2.7±1.0 %/Torr vs. CA: 4.0±1.1 %/Torr; P=0.01). Despite the small sample size, these data suggest that microvascular function is impaired in young otherwise healthy AAs.
Effects of modulators of AMPK on TASK K+ channel and [Ca2+] in rat carotid body glomus cells
Authors: Kim Donghee , Kang Dawon , Martin Elizabeth A , Kim Insook , Carroll John L
Published: June 2021

Acute hypoxia depolarizes and elevates intracellular Ca2+ concentration ([Ca2+]i) in carotid body glomus cells. Recent studies suggest that AMP‐activated protein kinase (AMPK) mediates these effects of hypoxia by inhibiting background K+ channels such as TASK. To obtain additional evidence for the role of AMPK, we studied the effect of modulators of AMPK on TASK activity in cell‐attached patches of rat glomus cells. Perfusion of glomus cells for ~10 min with AICAR (1 mM) and A769662 (0.1 mM), two activators of AMPK, did not inhibit TASK activity or elicit depolarization. When cells were incubated with AICAR (1 mM), A769662 (0.1 mM), or DMSO (0.1%) for 2–3 hr, TASK activities in the three groups were also not significantly different. Furthermore, hypoxia inhibited TASK activity by 72% and 75% in cells pretreated with AICAR or A769662, respectively. Both AICAR and A769662 failed to increase [Ca2+]i in glomus cells. Compound C (40 μM) did not prevent hypoxia‐induced inhibition of TASK. AICAR and A769662 phosphorylated AMPKα in PC12 cells, and this was prevented by Compound C. Together, these findings suggest that inhibition of TASK by hypoxia is not mediated by AMPK. (supported by NIH and AHA)
In vitro antibacterial activity of the leucocyte and platelet‐rich fibrin clot exudate (L‐PRF)
Authors: Bozelli Ana Flavia , de Menezes Reiff Rodrigo Bezerra , Semeghini Dalmyr Osmar , Corbi Pedro Paulo , Lustri Wilton Rogério , Amaral André Capaldo
Published: June 2021

The leucocyte and platelet‐rich fibrin (L‐PRF) is a potential tool to investigate the effects attributed to platelet concentrates. Although able to stimulate tissue repair, there are no reports regarding its antibacterial activity. To evaluate the action of the exudate extracted from the L‐PRF in vitro, blood samples of 10 male volunteers (ages 18–24) were centrifuged in clot activator tubes. The clots were shaken vigorously after 1 and 6 hours of activation for the extraction of exudate. Platelet (PLA), leucocyte (LEU) and erythrocyte (ERY) counting was performed in whole blood, platelet‐poor serum (PPS) and exudate. The antibacterial activity was investigated by disk diffusion technique in strains of Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (27853) and Staphylococcus aureus (25923), using specific antibiotics as positive controls. Cell counting in whole blood yielded a value of 6 ± 1 × 103 for LEU, 226 ± 28 × 103 for PLA and 5 ± 0.3×106 for ERY; 1‐hour PPS: 0.1 ± 0.1 to LEU and 0.5 ± 0.8 for PLA; 6‐hour PPS: 0.1 ± 0.2 to LEU and 0.4 ± 0.7 for PLA; exudate 1 hour: 5 ± 4 to LEU and 82 ± 56 for PLA; exudate 6 hours: 8 ± 7 to LEU and 211 ± 240 for PLA. The values of the ERY in the exudate and PPS were negligible. The exudates and PPS samples did not generate an inhibition zone and the positive controls exhibited halos with specific inhibition patterns. The L‐PRF exudate showed no antibacterial activity.

Financial Support: CNPq Brazil
Determination of the Glycemic Index of a Weight Loss Meal Plan Program Using Energy Controlled, Pre‐packaged Products
Authors: Nichols Meghan , Jenkins Alexandra M , Fabricatore Anthony N , Vuksan Vladimir , Wolever Thomas MS , Daggy Bruce P
Published: June 2021

Background

Consumption of low glycemic index (GI) diets has been associated with a range of health benefits including weight loss. Meal plans incorporating pre‐packaged, energy‐controlled, low‐GI products may therefore be a convenient and effective intervention for weight management. However, GI is affected by the degree of food processing. This suggests that a low‐GI diet based on such foods could be difficult to construct.

Objective

Determine the GI of a meal plan based on a line of prepackaged foods designed to have a low‐GI.

Methods

Sixty‐six foods, including 16 breakfast, 15 lunch, 13 dinner and 22 snack items, were tested. Groups of ten healthy subjects tested each food once and a control food three times. The GI of test foods was calculated using standard methodology from capillary blood samples obtained at −5, 0, 15, 30, 45, 60, 90, and 120 minutes. GI for the full meal plan was estimated from tested foods and recommended supplemental grocery items.

Results

49 foods fell in the low GI range, 13 in the medium and 4 high. Estimated dietary GI of the full meal plan was in the low range (GI = 44).

Conclusion

This study demonstrated that an energy‐controlled, low‐GI diet can be assembled using a line of pre‐packaged foods, thus providing an option in designing research or clinical interventions requiring a low‐GI diet.

Funding Source: Nutrisystem, Inc.
Over‐expression of RAGE by proximal lung epithelial cells causes an inflammatory response in adult mice
Authors: Bennion Brock G , Bodine B Garrett , Leatham Emma , Wright Alex J , Jergensen Zac R , Erickson Connor J , Jones Cameron M , Johnson Jeff P , Knapp Steven M , Reynolds Paul R
Published: June 2021

Receptors for advanced glycation end‐products (RAGE) are multiligand cell‐surface receptors expressed abundantly by pulmonary epithelium. Our lab has discovered RAGE‐mediated effects in lung inflammation induced by tobacco smoke and environmental pollutants; however, the precise contribution of RAGE to proximal airway inflammation was yet unknown. We generated a Tet‐inducible transgenic mouse that conditionally over‐expresses RAGE using the Clara Cell Secretory Protein (CCSP) promoter active in proximal airway Clara cells. RAGE was induced for 40 days from weaning (20 days of age) until sacrifice date at 60 days. Compared to controls, immunohistochemistry, immunoblotting, and qPCR revealed significant RAGE up‐regulation; however, H&E staining revealed no detectible morphological disturbances. Freshly procured bronchoalveolar lavage fluid (BALF) from RAGE TG mice had significantly greater total protein content, total leukocyte quantity, PMN abundance, and cytokine concentration compared to age‐matched control littermates. These data support the concept that RAGE up‐regulation in lung airways may participate in inflammation of the proximal airway. Supported by the Flight Attendant's Medical Research Institute (FAMRI, PRR) and a BYU MEG (PRR).
BrightMed: an opportunity to open up anatomy to the next generation
Authors: Evans Darrell John Rhys , Beaney Darren John
Published: June 2021

BrightMed is a dedicated widening participation program created within Brighton and Sussex Medical School targeting gifted and talented pupils attending state schools in the region. Students are recruited in year 8 and begin the program in year 9. Over the next four academic years, students attend a number of activity days plus a week‐long summer residential. Currently BrightMed is working with 227 students. Anatomy features as a strong theme within the programme with sessions aimed at exploring the human body in health and disease. Highlights include living anatomy sessions, dissection and interactive ‘Anatomy Prof’ sessions. To date, all students who have applied to study medicine and have met the academic criteria have had at least one interview and of those 80% have received offers. The success of the programme has recently been recognised nationally and features as a best practice case study in a report for the UK Government.
Is salamander limb regeneration really perfect? First detailed anatomical and morphogenetic analysis of forelimb muscle regeneration in GFP‐transgenic axolotls as a basis for regenerative, developmental and evolutionary studies
Authors: Diogo Rui , Tanaka Elly
Published: June 2021

The axolotl Ambystoma mexicanum is one of the most used model organisms in evolutionary, developmental and regenerative studies, particularly because it is commonly said that it can reconstitute a completely normal and functional fore/hindlimb after amputation. Surprisingly, there is not a publication that tested the hypothesis that each regenerated forelimb muscle is really a ‘perfect’ copy of the original muscle. Here we describe the regeneration of the arm, forearm, hand, and some pectoral muscles in transgenic axolotls that express GFP in muscle fibers and report that: 1) there is a high percentage (43%) of muscle anomalies in the regenerated forelimbs; 2) however, on average in each regenerated forelimb there are anomalies in only 2.5% of the total number of muscles; 3) one of the most notable and common anomalies (in 35% of the regenerated forelimbs) is the presence of a fleshy coracoradialis at the level of the arm; this is a particularly remarkable configuration because in axolotls and in urodeles in general this muscle only has a thin tendon at the level of the arm; 4) during forelimb muscle regeneration there is a clear proximo‐distal and radio‐ulnar morphogenetic gradient but also a ventro‐dorsal gradient, which to our knowledge was not previously reported in the literature. We discuss the broader implications of these observations for regenerative, evolutionary, developmental and morphogenetic studies.

Grant Funding Source: Start‐up package of Howard University College of Medicine
Sources of dietary guidance‐based food groups and empty calories among the U.S. population in relation to income and race/ethnicity
Authors: Kirkpatrick Sharon I , Reedy Jill , Krebs‐Smith Susan M
Published: June 2021

Variation in rates of adherence to recommendations for dietary guidance‐based food groups have been observed in relation to socioeconomic status. In this study, we examined top sources of food groups and empty calories among the U.S. population (aged ≥2 years) by income and race/ethnicity using dietary recall data from the 2003–2004 National Health and Nutrition Examination Survey (n=8272). Income groups were identified using the poverty income ratio (≤130%, 131–185%, ≥186%) and race/ethnic groups included non‐Hispanic white, non‐Hispanic black, and Mexican‐American. The top contributors to many food groups were strikingly similar across income and race/ethnic groups; however, some exceptions were apparent. White potatoes were the top source of vegetables for all income and race/ethnic groups with the exception of Mexican‐Americans, whose top source was beans. For non‐Hispanic Blacks, Mexican‐Americans, and the lowest income group, the top source of dairy foods was whole milk whereas it was reduced fat milk for non‐Hispanic whites and the upper two income groups. Soda, energy and sports drinks were the top source of added sugars for all groups, whereas the top sources of solid fats differed. Understanding variations in sources of food groups and empty calories among the population may assist in better targeting individual‐ and environmental‐level interventions to improve dietary intakes.
Using models and modeling to interpret science to students, teachers and families
Authors: Herman Tim , Colton Shannon , Franzen Margaret , Hoelzer Mark , Vogt Gina
Published: June 2021

The MSOE Center for BioMolecular Modeling (CBM) is an instructional materials development laboratory. Founded on a core technology of 3D printing, the CBM creates accurate physical models of proteins, based on the atomic coordinates of solved structures. These models have proven to be powerful “tools of engagement” in our science education outreach programs targeted to students, educators and the general public. A physical model of a protein functions as an external representation of the “mental model” that is carried around in the head of a scientist. As an external representation of the researcher's mental model, the physical model is a tangible artifact that can be shared by the expert (researcher) and the novice (student, teacher, parents) as it stimulates meaningful conversations about science, between people of disparate backgrounds and levels of experience. Physical models of proteins are supplemented with other hands‐on, active learning instructional tools and featured in two new professional development programs for high school teachers and their students; (i) Genes, Genomes and Personalized Medicine, and (ii) Drugs, Drug Targets and You. These same tangible instructional tools are featured in a series of Science Cafés designed to communicate current advances in the biomolecular sciences to the general public.
Impact of “En Balance” Culturally Sensitive Educational Program on Lifestyle Changes among Hispanics with Type‐2 Diabetes
Authors: Al Abdrabalnabi Ahmed A. , Marghalani Abdullah A. , Beeson Larry , Firek Anthony , Schulz Eloy , De Leon Marino , Balcazar Hector , Cordero‐MacIntyre Zaida R.
Published: June 2021

O

We assessed the effect of a culturally sensitive educational program on lifestyle changes among Hispanics with type‐2 diabetes mellitus (DM2).

M

Thirty‐four participants (25 females and 9 males) joined a three‐month education intervention program. Participants completed a Family Habits Frequency Questionnaire at baseline and after three months. Participants were Spanish speaking Hispanics with DM2 aged 37–69 years.

R

Participants showed significant improvement toward acquiring healthier behaviors. For instance, participants read food labels significantly more often after the 3 months compared with baseline (P=0.0002). Also, there was a significant increase of physical activity among mothers such as doing 30 minutes of physical activity (P=0.04) and increasing in walk (P=0.01). The mean change of HbA1c (P=<0.001), fasting plasma glucose (P=0.003), and cholesterol/HDL ratio (P=<0.001) decreased significantly and HDL level (P=<0.001) increased significantly after the educational program.

C

The culturally sensitive educational program among Hispanics with DM2 showed significant positive changes in participants’ lifestyle and behaviors.

Grant Funding Source: CMS 03–00335 Health Services Research and NIH award 5P20MD001632
Fifteen Years Of The Usda's National Food And Nutrient Analysis Program: Improving National Food Composition Data
Authors: Pehrsson Pamela , Haytowitz David
Published: June 2021

Since 1997, the National Food and Nutrient Analysis Program (NFNAP), a USDA‐NIH collaboration, has improved the data quantity and quality in USDA food composition databases through the analysis of nationally representative samples of foods and beverages. The focal database is the USDA National Nutrient Database for Standard Reference (SR), the authoritative food composition database in the US and the international research community. An updated sampling plan based on 2010 US Census data is being implemented to account for recent demographic changes. Foods are collected nationwide from retail locations, composited, and sent with QC materials to commercial labs/cooperators for analysis. Results, self‐weighted by market share, often show wide variation among retail brands, restaurant chains, and botanical varieties when compared to values of unknown/non‐representative sampling or to older data. Sodium in French fries, for example, varies widely among top restaurant chains sampled under NFNAP (46–521 mg/100g, p<.0001). As manufacturers reformulate their foods in response to public health concerns (e.g., trans fatty acids, sodium) it is imperative for food composition databases to keep current. Since its inception, 1440 foods (representing almost 100,000 data points) have been added or updated in SR; these data are critical to health researchers, nutrition policy makers, the food industry and consumers.
Lycopene up‐regulates mRNA expressions of SIRT and adiponectin and down‐regulates PPARγ and IL‐6 in adipose tissue of hypercaloric diet‐fed rats
Authors: Luvizotto Renata AM , Nascimento Andre F , Miranda Natália CM , Pierine Damiana T , Imaizumi Erika , Presti Paula T , Correa Camila R , Yeum Kyung‐Jin , Wang Xiang‐Dong , Ferreira Ana Lucia A
Published: June 2021

Objective

To evaluate the effect of lycopene supplementation on inflammation in adipose tissue of obese rats.

Methods

Male Wistar rats were first fed control diet (C, n=6) or hypercaloric diet (HD, n=12) for 6 wks. Afterwards, the HD rats were randomized into two groups: HD [placebo (corn oil)] or HD+L [lycopene (10 mg/Kg BW/day in corn oil)] by gavage for 6 wks. Plasma lycopene concentration was analyzed by a HPLC system. The mRNA expressions of SIRT1, FoxO1, adiponectin, PPARγ, MCP‐1 and IL‐6 in adipose tissue were determined by real time PCR.

Results

Lycopene was detected in plasma (24 nmol/L) from HD+L group only. The mRNA expressions of adiponectin, SIRT1 and FoxO1 decreased significantly but PPARγ, MCP‐1 and IL‐6 increased significantly in adipose tissue from HD group, as compared with those of C group. Although there was no difference in adiposity index between the HD and HD+L group, lycopene supplementation restored the reduced expressions of SIRT1 and FoxO1 to the levels of the C group and inhibited the up‐regulations of PPARγ, MCP‐1 and IL‐6 mRNA expressions in the HD+L group.

Conclusion

Lycopene modulates several key inflammatory genes in adipose tissue indicating its potential protective effect against obesity‐related inflammation.

Financial Support: FAPESP (2010/06100–9; 2010/19746–4; 2011/22786–0)
Evaluation of a Novel Method for Quantifying Nutrition Content in Medical Student Patient Write‐ups
Authors: Seres David Stuart , Bogart Jane S , Goglia Alexander G , Peever Steven M. J. , Marr Joshua R
Published: June 2021

Objective

To assess a novel scoring system for evaluating nutrition content in patient write‐ups performed by Columbia University medical students.

Methods

A binary scoring system (inclusion scored one point, omission zero) composed of 8 nutrition‐related criteria pertinent for inclusion in all patient assessments (e.g. height, weight, BMI, weight history, diet, supplements, etc) was created. To evaluate the scoring system, Nutrition Master's students separately read and scored sequential sets of five write‐ups, performing active analysis, revision, and creating rules, until a maximum one‐point discrepancy in the final score of a maximum of one out of five write‐ups was achieved. Two groups of two student raters each independently scored an identical discovery set of 52 write‐ups. Inter‐rater and cross‐group reliability were calculated.

Results

The rater's scores for the discovery set had an agreement, measured by multiple item inter‐rater reliability, of 0.999 in the first rater group and 1.000 in the second rater group. Amongst all raters, the average write‐up score variance was 0.194 and the average agreement was 0.999.

Conclusions

The current study serves to establish inter‐rater reliability for an objective method of assessing medical student patient write‐ups for essential nutrition‐related content. These scores will be tracked over years to determine the impact of curricular interventions.
Association between changes in plant protein and mineral intakes and blood pressure as part of a dietary portfolio: a randomized controlled trial
Authors: de Souza Russell Jude , Ireland Christopher , Pellini Christopher , Galange Patrick , Jenkins David JA
Published: June 2021

Background

Combining foods with lipid‐lowering properties (“dietary portfolio”) lowers blood pressure [BP].

Objective

To determine associations between dietary portfolio components and BP.

Methods

Parallel substudy (n=97) with hyperlipidemia from 2 Canadian academic centers (Toronto and Vancouver). Participants received dietary advice on either a low‐saturated fat therapeutic diet (control) or a dietary portfolio emphasizing plant sterols, soy protein, viscous fibers, and nuts. Dietary portfolio counseling was delivered over 6 months at 2 intensities: “routine” (2 clinic visits); and “intensive” (7 visits).

Results

Mean systolic blood pressure [SBP] fell 2.7 mmHg (95% CI: 0.3 to 5.6) on the intensive portfolio (n=29), 4.4 mmHg (1.3, 6.4) on the routine portfolio (n=31), and 0.5 mmHg (−3.1, 2.0) on the control (n=37). Diastolic blood pressure [DBP] fell 2.0 (0.3 to 4.7), 2.3 (0.7 to 3.9), and 1.0 mmHg (−2.6 to 0.6), respectively. The dietary portfolio reduced SBP and DBP compared with control (P<=0.021). Increased plant protein intake (%) correlated with reductions in SBP (r=−0.224, p=0.027) and DBP (r=−0.238, p=0.019), as did increased magnesium (%) (SBP: r=−0.169, p=0.048; DBP: r=−0.243, p=0.004). Changes in potassium and sodium intakes were not associated with BP changes.

Conclusion

Higher plant protein and magnesium intakes were associated with favourable changes in BP. Primary funding: CIHR.
Branched Chain Fatty Acids (BCFA) concentrations in US foods and estimated BCFA intake in the US population
Authors: Ran‐Ressler Rinat Rivka , Bae SangEun , Lawrence Peter , Brenna James Thomas
Published: June 2021

Introduction

Branched chain fatty acids (BCFA) are a class of primarily saturated fatty acids (FA) with a methyl branch on the carbon chain. We recently showed that BCFA are constituents of the term newborn infant's gut, and feeding neonatal rats with a rat formula with BCFA of the type found in US food altered gut microbiota and reduced the incidence of Necrotizing Enterocolitis. Thus, BCFA may have beneficial effects on proper gut development and function.

Our aim was to analyze the levels and distribution of BCFA in common foods consumed in the US, and to estimate the amount of BCFA intake from these foods. Because BCFA have been reported in cow's milk and are a major component of bacterial membranes, we were specifically interested in BCFA from dairy‐based and fermented foods.

Methods

Food samples were purchased from local stores. FA analysis was carried out using routine methods.

Results

BCFA concentration in ruminant foods ranged from 1.4–2.7%w/w. Intake of BCFA per capita/d from foods was about 400mg/d. If Americans consumed the recommended amount of servings from the dairy food group, daily BCFA intake would increase.

Conclusions

This is the first report of BCFA concentrations in American foods. The consumption of BCFA per capita per day is higher than the consumption of bioactive n‐3 FA. The prominence of BCFA in the US food supply and their bioactivity strongly suggest that BCFA health effects should be studied.
Arachidonic acid and Docosahexaenoic acid attenuate inflammation in human fetal but not adult intestinal epithelial cells
Authors: Wijendran Vasuki , Zhu Weishu , Brenna J T , Innis Sheila M , Walker W Allan
Published: June 2021

Necrotizing enterocolitis in preterm infants is characterized by excessive inflammatory response of the immature gut. The objective of the current study is to investigate the effects of long chain polyunsaturated fatty acids docosahexaenoic acid (DHA) and arachidonic acid (AA), at physiological range of concentrations present in human milk, on inflammation in human fetal and adult intestinal cells and delineate their mechanisms of action. Human fetal and adult (Caco2) intestinal epithelial cells in culture were treated with 10, 50, 100 and 200uM concentrations of DHA, AA and palmitic acid (PA) conjugated to bovine serum albumin for 72hrs and challenged with IL1‐β. In human fetal enterocytes, AA suppressed IL‐8 and IL‐6 secretion in a dose dependent manner compared to control, DHA and PA. DHA at 200μM conc attenuated IL‐8 and IL‐6 secretion compared to control and PA. DHA enhanced fetal intestinal epithelial cell viability at 100 and 200μM conc compared to control, PA and AA. In contrast, in adult enterocytes, AA had a pro‐inflammatory effect with increased IL‐8 secretion and DHA did not show any dose‐ dependent effects. PA increased IL‐8 and IL‐6 secretion by fetal and adult enterocytes compared to DHA and AA. Thus, AA and DHA developmentally regulate intestinal inflammation and may protect preterm infants against necrotizing enterocolitis.

(Supported by a supplemental grant awarded to Dr. V. Wijendran, NICHD‐ R01‐ HD059126, PI: Dr. Walker)
Student understanding of glucose homeostasis: not so “sweet”
Authors: Wehrwein Erica A , Drobish Justin N
Published: June 2021

Undergraduate students exhibit a surprising lack of functional knowledge of glucose homeostasis. In addition, students are typically not aware of the clinical “gold standard” for assessing glucose homeostasis—the glucose clamp. Therefore, we have created an exercise focusing on these topics in our senior‐level Physiology Capstone Laboratory course at Michigan State University. This exercise is a “dry lab” that uses TA guided problem sets on: 1) hyperinsulimemic‐euglycemic clamp for normal and insulin resistant subjects, 2) hyperglycemic clamps for normal and type II diabetic subjects, and 3) hyperinsulimemic‐hypoglycemic clamps for normal subjects and those with impaired hypoglycemic counterregulation. Students work in small groups to complete graphs of controlled variables, measured variables, and predicted outcomes from each experimental scenario. The percentage of students earning a 4.0 on the pre‐lab assessment was 5.7% despite recently completing a full year of both Physiology and of Biochemistry lecture‐based coursework in this area, while 55.9% of students obtained a 4.0 on the post‐lab assessment (n=34). Further, the average score on the pre‐lab assessment was 59% (0.0 GPA), while the post‐lab assessment was 85% (3.5 GPA). In summary, this exercise greatly improves understanding of glucose homeostasis.
The role of keratinocyte‐derived α‐melanocyte‐stimulating hormone and endothelin 1 signaling in a murine melanoma‐susceptible model
Authors: Kyryachenko Sergiy , Schumann Canan , Indra Arup
Published: June 2021

Nuclear receptor RXRα is a member of steroid hormone retinoid‐X‐receptor family. Mice with epidermal deletion of RXRα (RXRαep−/−) develop melanoma‐like tumors upon chemical carcinogen treatment. This suggests that distinct non‐cell autonomous molecular events mediated by RXRα in keratinocytes may regulate melanoma progression.

Melanocyte homeostasis in skin is largely influenced by signals from neighboring keratinocytes, autocrine signals, and environmental factors, such as ultraviolet radiation (UVR). UVR affects melanocytes directly by damaging the DNA and indirectly by stimulating the synthesis of keratinocyte‐derived factors, such as endothelin 1 (EDN1) and α‐melanocyte stimulating hormone (α‐MSH).

Pharmacological inhibition of EDN1 signaling enhances UV‐induced DNA damage in melanocytes, but it does not affect the proliferation of keratinocytes, melanocytes, and dermal cells in control (RXRαL2/L2) and RXRαep−/− mice. In contrast, inhibition of α‐MSH signaling reduces UV‐induced DNA damage in all skin cell types. Moreover, it also leads to minimal increase in proliferation of dermal cells, but not of keratinocytes and melanocytes in the skin of RXRαL2/L2 and RXRαep−/− mice.

This research will help to evaluate strategies for preventing and/or curing UV‐induced melanoma by using a combination of antagonists and/or agonists of keratinocyte‐derived factors.

Grant Funding Source: NIEHS grant R01ES016629 (to AI)
Canonical Wnt/ β‐catenin Signaling Pathway mediates Shear Stress‐Activated Angiopoeitin‐2 expression and vasculogenesis
Authors: Yu Fei , Li Rongsong , Jen Nelson , Chi Neil , Lien Ching‐Ling , Hsiai Tzung
Published: June 2021

It is well known that fluid shear stress is intimately involved in vascular stability and differentiation. In particular, low or oscillatory shear stress (OSS) upregulates Angiopoietin‐2 (Ang‐2) in human aortic endothelia cells (HAEC) via VEGF‐dependent mechanism. We hypothesized that Wnt signaling is implicated in shear stress activated vasculogenesis via modulating Ang‐2. Activation of Wnt signaling was assessed with TOPflash lentivirus reporter. After 8 hours of oscillatory flow, HAECs infected with lentivirus of Wnt reporter demonstrated markedly increased luciferase activity compared to cells under static condition, suggesting an activation of canonical Wnt signaling pathway with OSS. We further demonstrated that Wnt3a, a Wnt activator, upregulated Ang‐2 mRNA expression, whereas DKK‐1, a Wnt inhibitor, inhibited Ang‐2 expression in both HAEC cell lines and Tg(hsp70:dkk1‐GFP) zebrafish embryos 72 hours post‐fertilization (hpf). Knock‐down of Ang‐2 by siRNA in HUVEC resulted in impaired tube formation. Similarly, Ang‐2 knock‐down with morpholino oligomers in 72 hpf Tg(flk1:GFP) zebrafish embryos caused impaired systematic vasculature development. Taken together, our findings suggest that Wnt/β‐catenin signaling pathway mediates shear stress‐activated endothelial Ang‐2 and regulates vasculogenesis. These studies were supported by NIH HL083015 and HD069305.

Grant Funding Source: NIH
Molecular organization of the endocannabinoid signaling system in the superficial spinal dorsal horn of rodents
Authors: Antal Miklos
Published: June 2021

Although it is generally accepted that the endocannabinoid system plays important roles in spinal pain processing, the molecular organization of the endocannabinoid signaling system in the superficial spinal dorsal horn is poorly defined that renders the interpretation of the effect of cannabinoids on pain processing spinal neural circuits dubious. Thus, using immunocytochemical methods at the light and electron microscopic levels, we investigated in the superficial spinal dorsal horn the cellular distribution of CB1‐Rs, as well as DGLá, NAPE‐PLD, enzymes synthesizing the two major endocannabinoids, 2‐AG and anandamide, respectively, as well as MGL, enzyme terminating the activity of 2‐AG.

Our results suggest that the activity dependent release of endocannabinoids activates a complex signaling mechanism in pain processing spinal neural circuits into which both neurons and glial cells may contribute. The contrasting distribution of CB1‐Rs and the enzymes synthesizing and degrading endogenous cannabinoids indicate that endocannabinoid signaling mechanisms may evoke both short and long lasting modulation of neural activities underlying pain processing in the superficial spinal dorsal horn.
Regulation of origin activation during Drosophila follicle cell gene amplification
Authors: Hua Brian , Li Sharon , Kim Jane C. , Orr‐Weaver Terry L.
Published: June 2021

We are utilizing the Drosophila follicle cell gene amplification system to better understand the mechanisms of DNA replication origin activation in metazoans. During gene amplification, specific origins of replication undergo repeated rounds of activation at six distinct sites in the follicle cell genome. These Drosophila Amplicons in Follicle Cells (DAFCs) utilize the same replication machinery as normal DNA replication, including the Origin Recognition Complex (ORC), DUP/Cdt1, and the MCM2–7 helicase. However, the mechanisms controlling the activation of amplification origins remain poorly understood. We identified an origin at DAFC‐62D whose activation is dependent upon transcription. We will present experiments utilizing transgenic DAFC‐62D transposon insertions to test whether transcription is required locally for origin activation. Additionally, we are investigating the effects of local chromatin on origin activation. We have identified cis‐acting position effects acting over at least 5 kb that repress ORC binding and amplification at DAFC‐22B.
A molecular analysis and clinical follow up: a strategy for hereditary cancer prevention
Authors: Soares Barbara Luisa , Santos Luciana Lara , Lima Karine Evangelista , Mayrink Luiz Henrique , Valentin Aline Angélica Souza , Oliveira Eneida Santos , Lemos Sara , Lopes Debora Oliveira , de Azevedo Barbosa Leandro , Rodrigues Angelica
Published: June 2021

According to the Brazilian National Institute, the incidence of cancer in Minas Gerais state, Brazil, is approximately 50.000 new cases per year. 5–10% of these cases are associated with hereditary predisposition. This project is a partnership among the public Cancer Hospital of Divinopolis, Federal University of São João del Rei and a Non Governamental Organization against Cancer. The objective is to structure a center for familial cancer control based in clinical and molecular analysis. Patients with clinical suspected familial cancer are assisted, familial history is collected and heredograms are made in Progeny software. Risk assessment is done and families who fill clinical criteria for hereditary cancer are advised about the importance of close follow‐up. After consent, 5 ml of blood are collected for molecular studies. Specific genes related to the suspected syndrome are screened for mutations. The techniques used are High Resolution Melting (HRM) and Sequencing. From January 2011 to date, 19 families were registered for different cancer familial syndromes. At this moment, different genes have been analyzed by HRM and are being sequenced. This study will help to identify the individuals with real risk, so that genetic counseling can be done. The partnership is a promissing solution for the establishment of clinical‐molecular projects in regions with limited economic resources. Support: CNPq and FAPEMIG.
Expanding the genetic code with pyroglutamate
Authors: Walsh Matthew , Benhaim Mark , Sheppard Kelly
Published: June 2021

Pyroglutamate (pGlu) forms via the cyclization of N‐terminal glutamine (Gln) or glutamate (Glu) residues in a wide‐range of proteins including amyloid β‐peptides associated with Alzheimer's disease and oconase, an anti‐cancer agent member of the RNaseA superfamily. To facilitate research into the role pGlu plays in these proteins, we are expanding the genetic code of E. coli to include the modified amino acid by reassigning the amber stop codon to pGlu. To include pGlu, we are modifying the archaeal RNA‐dependent Gln biosynthetic pathway [glutamyl‐tRNA synthetase (GluRS) and the amidotransferase GatDE] to synthesize pGlu on a mutant archaeal amber suppressor tRNA (tRNApGlu). The ability of the archaeal GluRS and mutant GatDE to use tRNApGlu as a substrate in vitro was investigated using gel‐shift assays and kinetic studies. The genes for tRNApGlu and the two aforementioned enzymes were cloned into a pRSF‐Duet vector and transformed into E. coli to determine if they can form pGlu‐tRNApGluin vivo. Read‐through of an amber codon in a genetic message is being verified using an enhanced yellow fluorescence protein (eYFP) reporter system. Site‐specific incorporation of pGlu into the protein will be validated by mass‐spectrometry. Once established, we will use the system to test the role of the amino acid in RNaseA.