bluuebunny/crossref_metadata_2025 huggingface dataset:
Mesenchymal Stem Cells treatment improves the endometriosis proliferation in cell culture
Authors: Goncalves Giovana , Invitti Adriana , Parreira Rafael , Schor Eduardo , Silva Ismael Dale Cotrim Guerreiro , Girão Manoel João Batista Castello
Published: June 2021

Endometriosis is a chronic gynecological disorder defined as the presence of endometrial tissue within extra‐uterine sites. The proliferation of topic and ectopic endometrial tissue was demonstrated in several trials. In endometriotic tissues and cultured endometriotic cells the expression profile of cell cycle and inflammatory proteins are modified, particularly p27 protein – reduced expression – and IL‐1β – increased expression –, both leading to an increased cell proliferation. The therapeutic use of an adenoviral p27 super‐expression vector (Adp27EGFP) in endometriotic cell cultures induces the cells back to normal proliferative levels. In order to improve the therapeutic potential of this gene therapy we combined it with cell therapy using human umbilical cord mesenchymal stem cells (hUCMSC). The levels of IL‐1β were evaluated in the culture medium of normal and endometriotic endometrial cells transfected or not with Adp27EGFP and Adnull, and treated with hUCMSC. Treatment with hUCMSC leads to increased IL‐1β expression reaching endometriotic levels after 14 days of treatment even in the normal endometrial cells. The therapeutic effects of Adp27EGFP were totally inhibited in the endometriotic cells treated with hUCMSC. These results suggest that the mesenchymal stem cells can have a role in the progression of the endometriosis and cannot be used to treat the disease.
Etoposide Catechol Is an Oxidizable Topoisomerase II Poison
Authors: Deweese Joseph E. , Jacob David A. , Gibson Elizabeth G. , Mercer Susan L.
Published: June 2021

Topoisomerase II is an essential nuclear enzyme involved in DNA metabolism. Anticancer drugs capitalize on the potentially genotoxic catalytic mechanism of topoisomerase II in order to cause permanent strand breaks. The anticancer agent etoposide prevents topoisomerase II from ligating cleaved DNA strands resulting in strand breaks and cell death. Surviving cells that attempt to repair this damage can incur translocations, which are associated with therapy‐induced secondary leukemias. Etoposide is metabolized into active catechol and quinone metabolites, but the contribution of these compounds to mutagenesis is unclear. We assayed etoposide catechol for the ability to poison human topoisomerase II. The catechol induces cleavage levels and double‐strand: single‐strand break ratio similar to those seen with etoposide. The catechol does inhibit ligation but does not inactivate the enzyme as the quinone does. However, the catechol can be oxidized using potassium‐ferricyanite to the quinone form and displays much higher potency and induces a higher double‐strand: single‐strand break ratio than observed with etoposide. We propose that the catechol can be oxidized to the quinone in cells causing high levels of double‐strand breaks, which may contribute to leukemogenic translocations. Research was supported by Lipscomb University College of Pharmacy Pharmaceutical Sciences Summer Research Program.
Physical and functional association of lactate dehydrogenase with mitochondria in intact and permeabilized skeletal muscle fibers
Authors: Kane Daniel A. , White Adrienne E. , Elustondo Pia , Hughes Meghan C. , Brebner Karen , Pavlov Evgeny
Published: June 2021

To clarify whether mitochondria oxidize lactate, mitochondrial respiratory oxygen flux (JO2) was measured during the sequential addition of various substrates and cofactors onto permeabilized rat red (RG) and white (WG) gastrocnemius muscle fibers, as well as isolated mitochondrial subpopulations. Addition of lactate did not alter JO2 in either RG or WG. However, subsequent addition of NAD+ significantly increased JO2, and was abolished by the inhibitor of mitochondrial pyruvate transport, α‐hydroxycinnamate. In parallel experiments with subsarcolemmal (SS) and intramyofibrillar (IMF) mitochondria isolated from RG, only SS exhibited significant NAD+ ‐ dependent lactate oxidation. To determine whether lactate dehydrogenase (LDH) is physically associated with mitochondria in RG, confocal microscopy was used to detect immunohistochemically stained LDH and mitochondrial proteins. LDH clearly colocalized with mitochondria in intact fibers. In permeabilized fibers, most of the LDH signal was lost in IMF but not SS regions. The LDH signal was absent in SS regions following trypsin treatment; however, colocalization was augmented in both SS and IMF regions with exogenous LDH + washing, suggesting LDH preferentially associates with muscle mitochondria. Collectively, these results suggest that LDH is strategically positioned within skeletal muscle fibers to functionally interact with mitochondria.
Muscle‐specific VEGF knockout disrupts thermoregulation without altering mitochondrial morphometry or activity
Authors: Mandler William Kyle , Audet Gerald N , Olfert I Mark
Published: June 2021

VEGF is believed to enhance thermogenesis in response to cold exposure through promoting angiogenesis, enhancing metabolism, and upregulating mitochondrial biogenesis. Using muscle‐specific VEGF gene deleted mice, we tested the hypothesis that loss of VEGF in skeletal muscle will affect thermoregulation.

VEGF knockout (KO) and wild‐type (WT) mice were exposed to 4°C for 6 hours. Core temperature was measured in 15 minute increments. Flow cytometry analysis was performed on isolated subsarcolemmal (SSM) and interfibrillar (IFM) mitochondrial subpopulations taken from cardiac and triceps surae muscle tissue of KO or WT to determine mitochondrial size and complexity.

Initial temperature was lower in the KO. During cold exposure, temperatures dropped more quickly in the KO group. The average time to reach a core temperature decrease of 8°C was shorter in the KO group. There were no differences in skeletal muscle SSM or IFM in size or complexity. In the cardiac tissue, both SSM and IFM exhibited greater size in the KO group. There were no significant differences in mitochondrial density for any tissue or subpopulation.

Muscle‐specific VEGF KO mice are unable to maintain core temperature to the same degree as WT in the face of a 4°C challenge and have a lower core temperature at room temperature. Skeletal muscle mitochondrial density and morphology does not appear to play a role in this effect.
Molecular mechanisms underlying aflatoxin‐induced mutagenesis
Authors: Lin Ying‐Chih , Li Liang , Burgers Peter M. , Stone Michael P. , Lloyd R. Stephen
Published: June 2021

Abstract Aflatoxin B1 (AFB1) is a known mutagen and carcinogen that is associated with hepatocellular carcinoma (HCC). Although the underlying mechanisms contributing to these events are far from complete, our investigations provide insight into the molecular mechanisms of aflatoxin‐accelerated carcinogenesis. Site‐specific mutagenesis assays in primate cells were used to examine the mutagenic potential of the primary and persistent AFB1‐DNA adducts, AFB1‐N7‐Gua and AFB1‐FAPY‐Gua, respectively. Both AFB1‐N7‐Gua and AFB1‐FAPY were highly mutagenic at frequencies of 45% and 97%, respectively, with the predominant base substitution being G to T transversions, a mutation that is consistent with observations made in aflatoxin‐associated HCC samples. Moreover, we present the first biochemical evidence that the replicative polymerase δ and the specialized translesion synthesis (TLS) polymerase ξ4 could bypass AFB1‐N7‐Gua accurately, while the other two TLS polymerases κ and ξ were responsible for G to T mutations that were induced by AFB1‐N7‐Gua and AFB1‐FAPY.

This work was supported by NIH R01 CA055678.
The effects of exercise under hypoxia on cognitive function
Authors: Ando Soichi , Hatamoto Yoichi , Sudo Mizuki , Kiyonaga Akira , Tanaka Hiroaki , Higaki Yasuki
Published: June 2021

Increasing evidence suggests that cognitive function improves during a single bout of moderate exercise. In contrast, exercise under hypoxia may compromise the availability of oxygen. Given that brain function and tissue integrity are dependent on a continuous and sufficient oxygen supply, exercise under hypoxia may impair cognitive function. However, it remains unclear how exercise under hypoxia affects cognitive function. The purpose of this study was to examine the effects of exercise under different levels of hypoxia on cognitive function. Twelve participants performed a cognitive task at rest and during exercise at various fractions of inspired oxygen (FIO2: 0.209, 0.18, and 0.15). Exercise intensity corresponded to 60% of peak oxygen uptake under normoxia. The participants performed a Go/No‐Go task requiring executive control. Cognitive function was evaluated using the speed of response (reaction time) and response accuracy. We monitored pulse oximetric saturation (SpO2) and cerebral oxygenation to assess oxygen availability. SpO2 and cerebral oxygenation progressively decreased during exercise as the FIO2 level decreased. However, the reaction time in the Go‐trial significantly decreased during moderate exercise under normoxia and hypoxia. Hypoxia did not affect reaction time. Neither exercise nor difference in FIO2 level affected response accuracy. These results suggest that acute exercise improves cognitive function, and that hypoxia has no effects on cognitive function at least under the present experimental condition. We received no financial support for this study.
Expression of Active and Latent Myostatin in Overload‐ Induced Hypertrophied Rat Skeletal Muscle
Authors: Affleck Paige A , Parcell Allen C , Thomson David M , Mack Gary W
Published: June 2021

Myostatin (GDF‐8) is the chief chalone in skeletal muscle and negatively controls adult skeletal muscle growth. The role of myostatin during overload‐induced hypertrophy of adult muscle is unclear. We tested the hypothesis that overloaded adult rodent skeletal muscle would reduce expression of myostatin. Overload‐induced hypertrophy was induced by unilateral tenotomy of the gastrocnemius tendon in 11 male adult Sprague‐Dawley rats followed by a 2‐week period of compensatory overload of the plantaris and soleus muscles. Western blot analysis was performed to evaluate changes in active and latent complex myostatin protein expression. Significant hypertrophy was noted in the soleus (494 ± 29 vs 405 ± 15 mg, p<0.05) and plantaris (289 ± 12 vs 179 ± 37 mg, p<0.05) muscles following overload. Overloaded plantaris muscle decreased expression of the active myostatin protein by 28.0 ± 6.6 % (p<0.01) while the myostatin precursor decreased slightly (p=0.04). Overloaded soleus muscle decreased expression of the active myostatin protein by 22.5 ± 5.4% (p<0.01) while myostatin precursor was unchanged. Myostatin latent complex expression decreased in the overloaded soleus by 11.4 ± 4.2% (p = 0.03) but was unchanged in the plantaris. These data support the hypothesis that the myostatin‐signaling pathway in overloaded muscles is generally reduced and contributes to muscle hypertrophy.
Matrix Metallopeptidases (MMPs) Expression Up‐Regulated In Peripheral Blood Mononuclear Cells Following Resistance Exercise
Authors: Carlson Lara A
Published: June 2021

Matrix metallopeptidases (MMPs) are proteolytic enzymes essential for degradation and remodeling of the extracellular matrix components (EMC), and are thought to participate in post‐exercise physiological adaptations. Recent evidence suggests that an acute bout of resistance exercise (RE) can produce muscle damage and local inflammation followed by tissue repair and remodeling. DNA microarrays have been utilized to examine the effects of exercise on global gene expression in human peripheral mononuclear cells (PBMCs). The present study utilized an acute bout of RE to examine the expression of inflammatory genes in PBMCs. Ten males performed an acute bout of RE utilizing the major muscle groups in the lower extremities. Venous blood was sampled at rest, post‐exercise, and at 2 hr post‐exercise, and was analyzed for gene expression using Affymetrix Genechip Arrays. Using a conservative false discovery rate of p<0.05 and a two‐fold change of or higher threshold, microarray analysis indicated that the greatest gene response was seen at the 2 hour post‐exercise. Immediately post‐exercise and at the 2 hr recovery time points, matrix metalloproteinase (MMP) 9, MMP 14, MMP 17, MMP 19, and MMP 25 showed significant up regulation. In conclusion, RE is capable of inducing several MMPs suggesting that they may play an important role in muscle remodeling following RET.
Submaximal cycling exercise stimulates mTOR signaling pathway in human skeletal muscle
Authors: Kakigi Ryo , Naito Hisashi , Yoshihara Toshinori , Ozaki Hayao , Kobayashi Hiroyuki , Okada Takao
Published: June 2021

It is known that eccentric muscle contraction promotes muscle protein synthesis. However, effects of concentric contraction on muscle protein synthesis are not clear. We determined whether high repetition of concentric contraction by cycling exercise stimulates mTOR signaling pathway in human skeletal muscle. Six young subjects (21±1 years) performed 90% VO2max cycling exercise at 90rpm for 5 minutes on bicycle ergometer. Muscle biopsies (vastus lateralis) were obtained before exercise and, at 1 and 3 h post‐exercise. mTOR signaling and markers of autophagy (LC3BI and LC3BII protein) were analyzed by using western blotting. Plasma GH, IGF‐1 and insulin levels significantly increased in post‐exercise period (p<0.05). Akt phosphorylation, upstream of mTOR, significantly increased at 1 h post‐exercise (p<0.05). S6K1 phosphorylation, a marker of mTOR activity, significantly increased at 3 h post‐exercise (p<0.05). At the same time point, LC3BII to I ratio were significantly decreased (p<0.05). These data suggest that high repetition of concentric contraction such as cycling exercise can stimulate mTOR signaling in human skeletal muscle, which might leads to promotion of protein synthesis and attenuation of autophagic protein degradation.

This study was partly supported by the Sasakawa Scientific Research Grant from the Japan Science Society, and a Grant‐in‐Aid for Young Scientists (B) No. 24700703.
Intramuscular and interstitial nitric oxide concentrations following doxorubicin administration in nude mice.
Authors: Ryan Talia , Fabris Sergio , MacLean David A
Published: June 2021

The purpose of the present study was to examine intramuscular (IM) nitric oxide (NO) concentrations in the gastrocnemius (G) and quadriceps (Q) muscles as well as interstitial NO levels in the G of immunodeficient nude mice (n=6) following the administration of doxorubicin (10mg/kg). Muscles were collected at day 3 and 7 post injection and interstitial concentrations were determined using the microdialysis technique. Gastrocnemius IM concentrations of NO were increased (P<0.05) from control (626±18 μmol/kg dw) to 823 ±48 and 831±22 μmol/kg dw at day 3 and 7, respectively. Similarly, IM quadriceps concentrations of NO increased (P<0.05) from control (599±13 μmol/kg dw) to 738 ±18 and 756±46 μmol/kg dw at day 3 and 7, respectively. In contrast, G interstitial NO concentrations did not change (P>;0.05) from control values (14.0±2.5 μmol/l) for either collection period (day 3 = 16.4±1.7. day 7 = 15.7 ±1.6 μmol/l). These data clearly demonstrate that intramuscular NO concentrations are increased as a result of doxorubicin administration, and remain elevated for up to 7 days following initial dosing. Interestingly, doxorubicin administration did not alter interstitial NO concentrations suggesting that the effects of anthracyclines on stimulating NO production was isolated to intramuscular mechanisms. Overall, this study suggests that doxorubicin administration stimulates intramuscular NO production and may be a result of increased oxidative stress.

Supported by NSERC
A new role for plasma membrane phosphatidylinositol 4‐phosphate (PI4P)?
Authors: Hammond Gerry R , Balla Tamas
Published: June 2021

Many crucial functions of the plasma membrane (PM) rely on the minor inner leaflet lipid, phosphatidylinositol 4,5‐bisphosphate [PI(4,5)P2]. The major intermediate in PI(4,5)P2 synthesis, PI4P, seems paradoxically to be superfluous for maintenance or function of PI(4,5)P2. Inhibiting the enzyme that generates PM PI4P, PI 4‐kinase (PI4K) type III α (PI4KA), leads to depletion of PI(4,5)P2 levels only when phospholipase C (PLC) is activated, suggesting that other PI4K activities synthesize PI(4,5)P2 in resting cells. Here, we set out to test this hypothesis. Using a combination of live cell imaging with fluorescent PI4P and PI(4,5)P2 biosensors, we show that type II or type III β PI4K inhibitors lead to reductions in endosomal and Golgi‐localized PI4P pools, whereas only an inhibitor of PI4KA produces a reduction in PM PI4P. However, no combination of these agents led to substantial depletion of PM PI(4,5)P2. Use of radiolabelled tracers demonstrated that PI4KA inhibitor caused reduced absolute levels of PI4P with little change in PI(4,5)P2, despite a dramatic reduction in acute PI4P and PI(4,5)P2 synthesis. Therefore, PI(4,5)P2 levels are maintained when PI4KA is blocked not because there is an alternative route of synthesis, but because PI(4,5)P2 turnover simply stops. Consistently, in cells depleted of PM PI4P by a PI4KA inhibitor, forced degradation of PI(4,5)P2 with a 5‐phosphatase yields a pool of PI4P, which rapidly runs down due to the absence of PI4KA activity to maintain it. These results suggest a novel function for PM PI4P, which is as a metabolic “buffer” whose levels and turnover can be rapidly sensed and adjusted in order to maintain the functionally crucial PI(4,5)P2 pool. This work was supported by the intramural research program of the Eunice Kennedy Shriver National Institute for Child Health and Human Development, NIH.
Changes in exercise hyperpnea are more attributable to limb movement frequency than pedal loading
Authors: Caterini Jessica Elizabeth , Wells Greg , Duffin James
Published: June 2021

The significance of limb movement frequency in determining ventilation during exercise remains a contested topic in respiratory research. We used two sinusoidal exercise protocols on a cycle ergometer to separately assess the effects of limb movement frequency i.e. cadence (CP) and pedal load (LP) on ventilation. During the first test, cadence was altered sinusoidally while pedal load remained constant. In the second test, pedal load changed sinusoidally while cadence was constant. Variables were fitted to a sinusoid with parameters amplitude, mean, period, and phase angle lag. Five participants completed both protocols. Mean (SD) cadence amplitudes differed between protocols (CP: 23.5 (0.67) rpm, LP: 0.47 (0.3) rpm, p<0.001). The phase lag angles for ventilation also differed, with sinusoidal cadence significantly smaller than sinusoidal load (CP: 23.7 (12.7) deg., LP: 48.1 (14.6) deg., p= 0.023). However the mean ventilation was not different between tests (CP: 44.5 (3.9) L/min, LP: 46.8 (4.2) L/min, p=0.99). VO2 and VCO2 did not differ between tests; eliminating the possibility that changes in ventilatory response between protocols resulted from differences in metabolism. Therefore we concluded that changes in exercise hyperpnea are more attributable to limb movement frequency than pedal loading.

Funding provided by the Faculty of Kinesiology and Physical Education at the University of Toronto
Diindolylmethane Attenuates TGF β Mediated Human Lung Fibroblast Proliferation
Authors: Galam Lakshmi , Bandyopadhyay Sayantani , Martinez Osvaldo , Abuelenen Toaa , Castillo Annie , Parthasarathy Prasanna , Lagishetty Venu , Lockey Richard , Kolliputi Narasaiah Kolliputi
Published: June 2021

Objective

Idiopathic pulmonary fibrosis (IPF) is a progressive scarring disorder characterized by the deposition of extracellular matrix causing impaired gas exchange with excessive proliferation and apoptosis‐resistant state of lung fibroblasts. Therefore, inhibitors of fibroblast proliferation offer considerable therapeutic promise. Diindolylmethane (DIM) is found in cruciferous vegetables and has been known for its anti‐cancer and anti‐proliferative properties. Surprisingly, DIM has never been studied in pulmonary fibrosis. In the current study, we tested the effect of DIM on human lung fibroblast proliferation and apoptosis.

Methods

Human lung fibroblasts were treated with TGF β, a fibroblast proliferation inducer, in the presence or absence of DIM. Proliferation and apoptosis were assessed by using commercially available kits. To further understand the mechanism of DIM, cell lysates were analyzed for gene expression analysis using RT PCR. Protein expressions were determined by western blot analysis.

Results

Our results showed that DIM suppressed TGF β induced proliferation in human lung fibroblasts but not apoptosis. In addition, DIM induced MMP1 expression and suppressed TGF β induced alpha smooth muscle actin and thrombospondin‐2 gene expression. Interestingly, DIM is not influencing ADAM‐TS2 expression (a pro‐fibrotic gene) in human lung fibroblasts.

Conclusions

Our findings demonstrated that the natural dietary compound DIM attenuates TGF β mediated pro‐fibrotic effects through regulating of MMP‐1 and thrombospondin‐2 in human lung fibroblasts. These findings suggest that DIM may be a potential therapeutic target for pulmonary fibrosis.
A Multidimensional Undergraduate Program for Educational Outreach Training in Neurophysiology
Authors: Castillo Jason Thomas , Mantaring Kristine Jane , Dagstanyan David , Dumaguindin Amanda Nicole , Valdez Matthew Christopher , Jreije Karim Michael , Kim Jin Ju , Dominguez Janei Marie , Bobis Sheila , Ferreyro Milotn Giovanni , Giraldo Andrea Veronica , Zepeda Nancy Carolina , Hanna Andrew Isam , Ellis James Benice , Okusanya Ayodeji Gregorio , Khan Minhaz , Curras‐Collazo Margarita
Published: June 2021

Neuroscience undergraduates enrolled in a 10‐week course entitled “Educational Outreach Training in Neuroscience,” that taught them skills necessary for effective outreach pedagogy. First, students learned presentation and leadership skills and read articles relevant to science. Students were then separated into smaller groups and worked at individual stations within a flex classroom that contained multi‐media facilities. Each group chose a topic such as the somatic motor system, reward pathway to develop for a 3–4 hr educational program to be delivered to 2–3rd, 5–6th grade and high school students. Each curriculum included interactive components such as hands‐on activities, songs, skits and outdoor games. A larger scale event, Brain Day, planned during Brain Awareness Week, was the highlight of the course. The Brain Day Program included a 20 min lecture, Q & A session, theatrical skit, activity booths, and campus/research lab tours. Neuroscience graduate students assisted in developing and implementing skits. As a result of this course: 1) undergraduates gained a deeper knowledge of neurophysiology, acquired/polished workplace skills, 2) UCR undergraduate and graduate students formed partnerships with local school teachers that continue to yield new collaborations and 3) local high elementary and high school students increased their knowledge of neurophysiology.
Could a puzzle be useful in the learning of the cardiac cycle?
Authors: Costa Rafaela , Sanches Andrea , Marcondes Fernanda Klein
Published: June 2021

We've developed a puzzle about the cardiac cycle to be used in Physiology classes to an undergraduate course of Dentistry. This puzzle presents one table with 5 columns, which contain the cardiac cycle phase, the Atrial state, the Ventricular state, the Atrium ventricular valves and the Pulmonary and aortic valves. Pictures of the cardiac cycle phases and chips are provided in order to complete the table. The chips indicate: the cardiac cycle phases, the options relaxed or contracted, which can be used depending on the atrial and ventricular states, and the options open or closed concerning the cardiac valves. In groups, the students are requested to complete the table with the chips meanwhile the professors discuss with them why they have chosen each chip. In addition, the groups have to discuss some questions about the transmission of the electric stimulus through the heart, the opening mechanism of cardiac valves, the role of gap junctions and atrium ventricular delay on the transmission of action potential in the heart. In order to assess how the students have evaluated the use of this puzzle, they were invited to answer a questionnaire. 52 out of 80 students agreed to answer it. All the students have signaled that the puzzle was useful to the comprehension of the cardiac cycle and after this puzzle they were able to establish the relationship between the morphological characteristics of the cardiac muscle and the cardiac cycle.
P2X receptor‐dependent erythrocyte damage by α‐hemolysin from Escherichia coli triggers phagocytosis by THP‐1 cells
Authors: Fagerberg Steen K , Skals Marianne , Leipziger Jens , Praetorius Helle A.
Published: June 2021

The pore‐forming exotoxin α‐hemolysin from E. coli causes a significant volume reduction of human erythrocytes that precedes the ultimate swelling and lysis. This shrinkage results from activation of Ca2+‐sensitive K+ (KCa3.1) and Cl− channels (TMEM16A) and reduced functions of both of these channels potentiate the HlyA‐induced haemolysis. This means that Ca2+‐ dependent activation of KCa3.1 and TMEM16A protects the cells against early haemolysis. Simultaneous to the HlyA‐induced shrinkage the erythrocytes show increased exposure of phosphatidyl‐serine (PS) in the outer plasma membrane leaflet, which is known to be a keen trigger for phagocytosis.

We hypothesize that exposure to HlyA elicits removal of the damaged erythrocytes by phagocytic cells. Cultured THP‐1 cells as a model for erythrocytal phagocytosis was verified by a variety of methods including live cell imaging. We consistently found the HlyA to very potently trigger phagocytosis of erythrocytes by THP‐1 cells. The HlyA‐induced phagocytosis was prevented by inhibition of KCa3.1, which is known to reduce PS‐exposure in human erythrocytes exposed to both ionomycin and HlyA. Moreover, we show that the P2X receptor inhibition, which prevents the cell damages caused by HlyA, also reduced that HlyA‐induced PS‐exposure and phagocytosis. Based on these results, we propose that erythrocytes, damaged by HlyA‐insertion, are effectively cleared from the blood stream. This mechanism will potentially reduce the risk of intravascular hemolysis
In situ Quantification of Cellular Cholesterol using a Molecular Sensor
Authors: Stec Ewa , Afsari Hamid Samareh , Kurilova Svetlana , Cho Wonhwa
Published: June 2021

Cholesterol has been implicated in regulation of physical properties of the plasma membrane, but its direct involvement in cell regulation through specific interactions with cellular proteins has not been thoroughly investigated. Also, the quantitative information about its cellular spatiotemporal dynamics and its local concentration fluctuation under pathophysiological conditions is lacking. To apply our recently developed quantitative lipid imaging technology to in situ quantification of cholesterol, we prepared a panel of cholesterol‐specific sensors from various cholesterol binding proteins that have different dynamic ranges of cholesterol detection. In situ quantification of cholesterol in various mammalian cells using these sensors revealed new insight into the spatiotemporal dynamics and local fluctuation of cholesterol under various physiological conditions.
Effects of doxorubicin on the expression of apoptotic‐related genes in 22rv1 prostate cancer cells (1010.12)
Authors: Fong Andy , Clack Beatrice
Published: June 2021

Doxorubicin (DOX) is a common chemotherapy drug used to treat a wide variety of cancer types. The drug is known as an intercalating agent that binds to DNA, inhibiting biosynthesis and ultimately resulting in apoptosis, or programmed cell death, of treated cells. The exact molecular mechanisms are complex and remain unclear. This project aims to elucidate the sequence of events that occur in DOX treated cancer cells leading up to apoptosis using the 22RV1 human prostate carcinoma cell line. The cells were cultured in complete medium, synchronized through serum starvation, and treated with DOX and harvested at 4 hour intervals up to 24 hours. The treated cells at all different time points then underwent a series of tests to measure cell survival rate and caspase 3/7 activity, which correlates to levels of apoptotic activity. The total RNA from the remaining treated cells were then extracted. Reverse transcriptase was used to synthesize cDNA, which was used as template on RT² Profiler™ PCR Array Apoptosis plates. The expression of 84 apoptosis related genes were evaluated using qRT‐PCR at each of the different treatment times providing details as to the onset and progression of apoptosis in the 22RV1 prostate cancer cells over a 24 hour period of DOX exposure.
Food consumption of Indian adolescents in a globalizing world (1014.5)
Authors: Shaikh Nida , Patil Shailaja , Ramakrishnan Usha , Cunningham Solveig
Published: June 2021

Aim: Describe the patterns of food consumption among adolescents in urbanizing environment in Bijapur city, India.

Methods: A cross‐sectional, cluster design study of 399 school‐going adolescents, ages 12‐16 years, attending six middle schools in Bijapur. A 16 items Food Frequency Questionnaire was administered by an interviewer and weekly consumption of key food groups (d/wk) was calculated. T‐tests were applied to test for differences in mean food consumption between adolescent boys and girls.

Results: Most commonly consumed food groups were tea/coffee (5.5 ± 2.8 d/wk), milk and milk products (4. 98 ± 2.8 d/wk), vegetables (other than green leafy vegetables (GLV)) (4.7 ± 2.3 d/wk), and packaged foods (biscuits, chips, chocolates) (4.7 ± 2.5 d/wk). Girls reported significantly (p<0.05) higher consumption (d/wk) of packaged foods (5.1 vs. 4.5), non‐GLV vegetables (5.0 vs. 4.3), ready‐to‐eat foods (1.5 vs. 0.8), and added dietary fat to prepared food (3.0 vs. 2.1) compared to boys. Boys reported significantly (p<0.05) higher consumption (d/wk) of eggs (2.0 vs. 1.5) and street foods (samosa, bhajji) (1.5 vs 0.9).

Conclusions: The high energy‐dense packaged and street foods intake combined with low intakes of fruits and GLV among adolescents supports the presence of the nutrition transition in urbanizing regions of India that have implications for increased risk for non‐communicable diseases risks in adulthood.

Grant Funding Source: Supported by NICHD Grant 3D43 and NIH Grant 1 R25 TW009337‐01 HD065249‐03S1 and
Lactation intensity and maternal weight loss at two months postpartum in women with recent gestational diabetes mellitus (1017.9)
Authors: Gunderson Erica , Matias Susana , Dewey Kathryn
Published: June 2021

Objective: To evaluate the association between lactation intensity and maternal weight loss at 2 months after gestational diabetes mellitus (GDM) delivery.

Methods: This analysis includes 960 participants (75% minority) in the Study of Women, Infant Feeding, and Type 2 Diabetes After GDM Pregnancy (SWIFT), who were diagnosed with GDM via Carpenter and Coustan criteria and delivered a live born infant of 37 weeks gestation or longer at Kaiser Permanente hospitals from 2008‐2011. We assessed lactation intensity since delivery, and estimated mean weight change (kg); defined as the difference between weight at delivery and weight at 2 months postpartum, among 5 lactation intensity groups using multiple linear regression models adjusted for pre‐pregnancy BMI, gestational weight gain, socio‐demographics and behaviors.

Results: Of 960, 25% and 34% were exclusively and mostly breastfeeding, respectively, and 15% were exclusively formula feeding. Mean (95%CI) weight losses were ‐1.4 kg (‐2.1, ‐0.6), and ‐1.2 kg (‐1.8,‐1.5) greater for exclusively and mostly breastfeeding groups versus the exclusively formula feeding group (p<.001) in fully adjusted models.

Conclusions: Intensive lactation was associated with greater weight loss within 2 months of delivery independent of lifestyle behaviors and other risk factors. Lactation may be beneficial in lowering adiposity among women at high‐risk for type 2 diabetes.

Grant Funding Source: National Institute of Child Health and Human Development (NICHD), R01 HD050625, R01 HD050625‐03S1, a
An evaluation of restaurants in low‐income communities using the Nutrition Environment Measurement Survey for restaurants (NEMS‐R) (1019.8)
Authors: Leone Ryan , Hemmingson Kaitlyn , Remley Daniel , Zies Susan , Kattelmann Kendra , Li Yijing , Kidd Tandalayo
Published: June 2021

As part of a multi‐state adolescent obesity project, we examined if signage, promotion, availability, quality and price of healthy food choices differed amongst sit‐down (SD), fast food (FF), or other restaurant (OT; i.e. fast casual) classifications. Restaurants (n=49) in three states were identified for audit using the validated Nutrition Environment Measurement Survey for restaurants (NEMS‐R). NEMS‐R evaluates the signage and promotion of healthy food items as well as their availability, quality and price. The standard NEMS‐R scoring rubric was utilized; higher scores indicate greater healthfulness and availability of menu items (range= ‐27 to 63). Establishments were excluded if they were not open to the general public or served predominantly to patrons over 21 years. Thirty‐one restaurants (SD=10, FF=6, OT=15) were assessed. Mean scores for SD, FF, and OT were 9.1 ±3.1, 20.8 ±19.4 and 11.3 ±17.8, respectively (min= ‐7, max= 53). Nutrition information was available at 10% of SD, 50% of FF and 60% of OT. Healthy items were promoted at 0% of SD, 17% of FF and 13% of OT. Approximately 5% of SD meals were healthy compared to 10% and 12% of FF and OT, respectively. Our results were not statistically significant warranting further study with larger samples and more statistical power. These data were easy to collect and provide an area of focus for obesity prevention efforts.

Grant Funding Source: Supported by the Agriculture and Food Research Initiative (AFRI) grant #2012‐68001‐19619 from the US
Impact of a point system program in increasing fruit and vegetable consumption among university employees (1021.12)
Authors: Fernandes Nicolle , Driver Stacey , Murr Rhonda , Friesen Carol
Published: June 2021

The purpose of this pilot study was to determine the effectiveness of “Colorful choices,™” a competitive, incentive‐based program aimed at improving fruit and vegetable consumption over a 4‐week period of time. Employees at a mid‐western university self‐enrolled in teams from their academic unit. Scoring was based on the team’s point average. Points were allocated as follows: vegetable servings = 2 points, fruit servings = 1 point and “Top Choice” selections (i.e., produce that may offer extra health benefits) = 1 bonus point. Point values were halved if a half‐serving was consumed. Electronic logging forms, team spreadsheets for leaders, and weekly e‐mails with recipes and tips were circulated to all team members. Although 170 participants enrolled in 18 teams, only 63% (n=107) completed the program; 10% (n=17) enrolled but never began, 10% (n=17) each dropped out after weeks 1 and 2, and 7% (n=11) after week 3. Repeated measures ANOVA revealed a significant increase in fruit and vegetable intake between the first week and the remaining three weeks, respectively (63±4 vs.77±5, 77±5, 78±5, p=0.001). Minor gender differences were noted (F= 89, M=18); females scored higher at the end of week‐1, however, males consistently scored higher for the remainder of the program (NS). The aggregate consumption was 9,919.5 servings of vegetable (equivalent to nearly 4,000 heads of lettuce) and 8,090.5 servings of fruit (equivalent to 2,700 pounds of apples) for a total consumption of 18,010 servings of produce. Based on participant feedback, 92% (n=22) found the weekly e‐mails beneficial and 83% (n=20) made use of the recipes and tips provided. Overall, 88% (n=21) indicated that the program helped to motivate their intake of fruits and vegetables, with 96% (n=23) opting to have the program offered again. In conclusion, a simple point system coupled with motivational weekly e‐mails in a competitive environment may favorably influence fruit and vegetable consumption among university employees.

Grant Funding Source: Working Well, Health Enhancement Program, Ball State University
The role of ready‐to‐eat cereal in the diets of children in the WIC Program: results from NHANES 2001‐10 and USDA Food Patterns Equivalents Database (1021.2)
Authors: Albertson Ann , Joshi Nandan
Published: June 2021

This study examined nutrient intakes, food sources of nutrients and food group consumption of children 1‐5 years participating in the WIC program by ready‐to‐eat cereal (RTE) consumption pattern (n=1941). These children were classified as either a RTE‐ eaters (n=1069) or non‐RTE‐eater (n=872) based on day‐1 intakes from NHANES 2001‐10. USDA Food Patterns Equivalents Database (FPED) was used to examine milk and whole grain intakes. RTE‐eaters had significantly (p<0.05) higher intakes of most micronutrients, including those identified as nutrients of need in the Dietary Guidelines, including vitamins A, C, D, calcium and iron (p<.05). RTE‐eaters obtained a substantial percentage of daily micronutrients from RTE; thiamin (32%), niacin (33%), folic acid (50%), iron (48%) and vitamin A (25%). Average daily fluid milk intake among RTE eaters (432 g/day) was significantly higher than non‐RTE eaters (369 g/day). Twenty‐four percent of the milk consumption among RTE‐eaters was consumed with RTE cereals. RTE‐eaters also had significantly higher intakes of whole grain (0.5 servings) compared to non‐RTE eaters (0.3 servings) (P<0.0001). On average, RTE cereal was the largest contributor to WG intake for children participating in the WIC program (45%).
A mobile phone‐based physical activity recall using daily activity pictures: helping to recall intensity and duration of physical activity (1021.7)
Authors: Pope Benjamin , Gallaway Patrick , Flores Uvalilia , Turner Rachel , Hongu Nobuko
Published: June 2021

The goals of this study were to develop a mobile phone‐based physical activity (PA) recall using periodic photographs and to compare reported minutes of moderate‐intensity PA (MPA) per day using the International Physical Activity Questionnaire (IPAQ). Participants used their cell phone to take 3 pictures (in the morning, afternoon, and evening) per day of their daily activities for 3 days per week. Participants were given the IPAQ within a week, and a PA recall (their minutes and intensity level of PA) was then conducted by researcher using their 9 pictures saved in their cell phone. The difference between the IPAQ reported minutes and the MPA by PA recall at each of four time intervals: baseline, 8‐week, 3‐months and 6‐months were obtained. To determine whether the difference in reported minutes between IPAQ and PA recall varied over time or with BMI, generalized estimating equations were used to account for the repeated measurements. Forty‐four females and 13 males, age 23.1 ± 5.74(SD) years, baseline BMI 26.2 ± 6.36(SD) participated in the study. The mean differences in the minutes at each of the time points were 16.3, 14.4, 3.4, and 10.7 minutes, respectively. Neither BMI nor time was significantly associated with the difference between minutes reported in PA recall and the minutes reported in the IPAQ. The first generation of cell phone‐based PA recall using activity pictures is well accepted and may help to predict better estimates of PA.
My Plate visual mobile application device for college students’ food intake tracking (1022.10)
Authors: Teo Yi Min , Pimentel Teresa , Wong Siew Sun , Cluskey Mary , Dorbolo Jon , Dinsmore Mark , Flynn Rebecca
Published: June 2021

Objectives: Test the feasibility of a mobile application to track college students' food intake.

Materials and Methods: The mobile app used a MyPlate visual concept for food data entry with a built‐in serving size estimation guide. Each user was instructed to record food intake by tapping the corresponding food group on the visual plate for each serving consumed. Subjects photographed and described their meal type, place and time of consumption and satiety upon completion. Eleven students (36% female, aged 18‐21) having two 3‐day food records participated in this feasibility study in Feb‐Mar 2013, including one online survey and a focus group). Meal photos were compared to meal descriptions, food groups, and serving sizes.

Results: The mobile app was convenient to use however, meal photos were incomplete. Suggested improvements included: 1) increase number of functions (e.g. caloric intake display); 2) enable viewing and editing of past entries (e.g, to adjust for dates and times and add forgotten foods; 3) a photo‐tagging system to match photos to corresponding meal data; and 4) increase text length for meal description.

Conclusion: Using MyPlate visual for dietary tracking is a feasible approach for college students who were familiar with food recordkeeping.

Grant Funding Source: PacificSource Healthcare
Development of non‐invasive assessments on vitamin A status by using skin levels of carotenoids as biomarker ő India study (1022.9)
Authors: Xiong Maxwell , Winterholler Erik , Wipfel Mia , Patel Dinesh
Published: June 2021

BACKGROUND: Some 127 million preschool children are Vitamin A Deficient (VAD), which is about one‐quarter of all preschool children in high‐risk regions of the developing world. Globally, approximately 4.4 million preschool children have visible eye damage due to VAD. Annually, about 500,000 preschool children go blind from VAD, and about two‐thirds die within months of going blind. Close to 20 million pregnant women in developing countries are also VAD, of which about one‐third are clinically night‐blind. Nearly one‐half of these cases occur in India. Invasive methods are common way for the diagnosis of VAD, but are costly and inconvenient to developing countries. OBJECTIVE: This is a continuation study from our US study. Skin levels of carotenoids directly depend on the diets and life style; therefore the data from different countries are important for development of non‐invasive methods for vitamin A, specially Indian subjects who are vegetarians. METHODS: More than 200 village preschool children, pregnant women, school students and adult men from Gujarat, India were enrolled by a local hospital; all subjects received the non‐invasive measurements on their skin levels of carotenoids by using a reflectance scanner. The blood samples were also collected for the analysis of vitamin A content. RESULTS: The average skin levels of carotenoids from young children and pregnant women were within normal ranges. However, the results from high school students caught our attentions because the reflectance scores were very low and there are some concerns on the potential risks of VAD for this group. The assay from plasma samples was consistent with the non‐invasive measurements. CONCLUSION: The noninvasive reflectance spectroscopy method is feasible to measure the vitamin A concentration in the blood, but more data from other regions and countries are needed to further confirm the correlations.
Predictors of overweight or obesity in school age children in Shanghai,China (1024.4)
Authors: Xiong Jianjing , Must Aviva , Wang Xiangdong , Gu Zhengyi , Xu Wenyan , Zhang Fangfang
Published: June 2021

Background. Childhood obesity is increasing dramatically in China, with causes presumed to be complex and as yet poorly understood. Identifying modifiable behavior risk factors can inform important insights into obesity intervention in school‐age children in China.

Methods. We assessed the trend of overweight and obesity in school‐age children in Shanghai from 2009 to 2012, and examined the contribution of diet and physical activity to the risk of overweight/obesity. Eight hundred and seventeen subjects from seven elementary schools in the Zhabei District of Shanghai, China were selected using stratified cluster sampling. Weight and height were measured by trained community health workers. A self‐administered lifestyle survey assessing diet and physical activity was completed by subjects’ caregivers. Overweight was defined as BMI z‐score >1 and 蠄2, and obesity was defined as BMI z‐score >2, according to the WHO definitions.

Results. Six hundred and eight‐six (84.0%) subjects had complete information on weight, height and lifestyle measures for all years 2009‐2012. The prevalence of overweight/obesity increased from 33.5% in 2009 to 41.7% in 2012 (an absolute 24.5% increase over the four‐year study period). Low vegetable consumption (grams/day per Kg of weight) (β =0.06, 95%CI: 0.03‐0.09), low fruit consumption (times/week per Kg of weight) (β =1.99,95%CI: 1.15‐2.83) and high screen time (hours/day) (β =0.22, 95%CI: 0.04‐0.41) were significantly associated with a high BMI z‐score at the final follow‐up, after adjusting for sex, age, parents’ education level.

Conclusions. Obesity is a significant public health problem in school age children in Shanghai, China. Increasing vegetables and fruits intake and reducing sedentary behavior may represent promising targets for intervention in this population group.
The impact of nutritional status on morbidities in mechanically ventilated critically ill children in PICUs (1024.8)
Authors: Bechard Lori , Duggan Christopher , Touger‐Decker Riva , Parrott J , Rothpletz‐Puglia Pamela , Byham‐Gray Laura , Heyland Daren , Mehta Nilesh
Published: June 2021

Mechanically ventilated children in pediatric intensive care units (PICUs) are at risk for hospital‐acquired infections (HAI) and prolonged hospitalizations. We performed a multicenter, international cohort study to determine the unique contribution of nutritional status to clinical outcomes in mechanically ventilated children in PICUs (N=1622). Nutritional status was 17.9% underweight (BMI Z score < ‐2), 54.2% normal weight (BMI Z score > ‐2 and <1), 14.5% overweight (BMI Z score > 1 and < 2), and 13.4% obese (BMI Z score > 2). Prevalence of HAI, length of stay, and ventilator‐free days (VFD) were evaluated using multivariate analyses, controlling for diagnosis, admission type, PICU location and size. Compared to normal weight, risk for HAI was significantly higher in underweight (OR 1.79; 95% CI 1.16, 2.75; p=.008), overweight (OR 1.34; 95% CI 1.04, 1.73; p=.02), and obese children (OR 1.50; 95% CI 1.09, 2.05; p=.01). Hazard ratios for hospital discharge were significantly lower among underweight (HR 0.71; 95% CI 0.61, 0.84; p<.001) and obese (HR 0.81; 95% CI 0.68, 0.96; p=.02) compared to normal weight children. Underweight was associated with significantly fewer VFD than normal weight (p<.001), overweight (p<.001), and obesity (p=.03). Nutritional status is an important contributor to morbidities in the PICU. Future studies exploring outcomes related to nutritional status during PICU admissions are needed.

Grant Funding Source: Supported by the Jean Hankin Nutritional Epidemiology award from the Academy of Nutrition and Dietet