bluuebunny/crossref_metadata_2025 huggingface dataset:
Coordinate Action of the Circadian Clock Protein Per1 and the Mineralocorticoid Hormone Aldosterone
Authors: Gumz Michelle , Richards Jacob , Cheng Kit-Yan , All Sean , Jeffers Lauren
Published: June 2021

Many metabolic and physiological processes, including renal function and blood pressure, exhibit a circadian pattern of variation. The molecular mechanism underlying the regulation of these processes by the circadian clock is not understood. We have previously shown that the circadian clock protein Per1 positively regulates the expression of the rate limiting subunit of the renal epithelial sodium channel (αENaC); and negatively regulates the expression of Claudin 1 (Cldn1), a component of epithelial tight junctions; and endothelin‐1 (ET‐1), a blood pressure‐regulating peptide hormone. In the present study we found that Per1 also positively regulates Rh family C glycoprotein (RhCG), an ammonia transporter in the kidney. The sodium‐regulating hormone aldosterone has been well established to regulate the transcription of αENaC, and was also found to upregulate the levels of Cldn1, ET‐1, and RhCG mRNA. Co‐immunoprecipitation of Per1 from nuclear extracts obtained from mouse renal cortex showed an interaction between Per1 and the glucocorticoid receptor (GR), a nuclear receptor that can bind aldosterone. Taken together, these results suggest a coordinate regulation of several genes by Per1 and aldosterone.
Molecular Interactions in PriC‐mediated DNA Replication Restart
Authors: Wessel Sarah , Marceau Aimee H , Hu Kaifeng , Westler Milo , Markley John , Keck James L
Published: June 2021

DNA replication restart is an essential process in bacteria that reloads DNA replication complexes (replisomes) that have ejected prematurely from replication forks due to encounters with impassable DNA damage or protein complexes. In E. coli and related bacterial species, the “replication restart proteins” (PriA, PriB, PriC, DnaT, and Rep) catalyze this activity. The 20‐kDa PriC protein is unique among these proteins in that it can mediate in vitro replisome reloading in the absence of other replication restart proteins. Precisely how PriC recognizes abandoned DNA replication fork structures and directs replisome reloading is currently unknown. We have found that the single‐strand DNA‐binding protein (SSB) forms a direct complex with PriC that we hypothesize is important for directing PriC reloading activities to abandoned DNA replication forks. PriC variants that fail that to bind SSB (but that retain DNA binding) were identified to test this hypothesis. Consistent with an essential role for the PriC/SSB interaction, these variants cannot catalyze the first step of replication restart (loading the DnaB helicase onto an SSB‐bound synthetic replication fork) in vitro. On‐going experiments aim to determine the solution structure of PriC and to assess the importance of complex formation with SSB in PriC‐mediated DNA replication restart in vivo. Research supported by GM068061 and T32‐GM07215.
Mitochondrial sensitivity to ADP in permeabilized fibers changed as a function of cardiac muscle compartments: left/right atria and ventricles of sheep heart
Authors: Chapolard Mathilde , Vaillant Fanny , Jais Pierre , Haissaguerre Michel , Dos Santos Pierre , Diolez Philippe , Deschodt‐Arsac Veronique
Published: June 2021

Because of the balance between ATP supply and ATP demand, alterations in cardiac energetics potentially affect cardiac work and may explain pathologies. Neverthelesss energetics alterations in each compartment remain unknown. Since mitochondrial oxidative phosphorylation is the main process of energy supply, mitochondrial sensitivity to energetic intermediates (e.g. ADP) is here investigated in a compartment‐specific way.

KmADP were obtained in sheep permeabilized fibers of right and left ventricles and atria using O2 consumption measurements (OROBOROS Oxygraph‐2k), carried out in Mir05 with glutamate (5 mM) and malate (2 mM), at 25Â°C.

Basal respiration (V0) and maximal ADP‐stimulated respiration (Vmax) were similar among atria or ventricles (meanV0 = 33 Â± 3.9 and meanVmax = 72 pmol.sec−1.mg−1 dry wt). Interestingly, KmADP were significantly higher in atria, especially in right one, than in ventricles, that were consequently characterized by a 1.5 to 3‐fold higher sensitivity to ADP compared to left and right atria.

In beating heart, for a given energy state of the cell, mitochondria are more responsive in ventricles than in atria and more responsive in the left than in the right atrium. Interestingly mitochondrial responsiveness is likely in adequation with requested work in each chamber of the healthy heart and may be closely related to the ventricle ability to withstand problems of arrhythmias.
Characterization of the proteome of cytoplamic lipid droplets in enterocytes in response to dietary fat
Authors: D'Aquila Theresa , Sirohi Devika , Kuhn Richard J. , Buhman Kimberly K.
Published: June 2021

Cytoplasmic lipid droplets (CLDs) are present and dynamically regulated in enterocytes, the absorptive cells of the intestine, in response to dietary fat. CLDs have a neutral lipid core surrounded by a phospholipid monolayer with associated proteins that have diverse cellular functions. The objective of this study was to determine the proteome of CLDs in enterocytes following a high fat dietary challenge. We isolated enterocytes from C57BL6 mice, two hours after a 200 μl olive oil gavage. The CLD fraction was isolated via sucrose gradient ultracentrifugation and analyzed by HPLC MS/MS. We identified 280 proteins in the CLD fraction representing proteins involved in diverse cellular functions including lipid and glucose metabolism. Lipid metabolism proteins identified include: perilipin 3, abhydrolase domain containing 8, acyl CoA synthetase 5, intestinal fatty acid binding protein (fabp), liver fabp, apolipoprotein AIV, and phospholipase c. Glucose metabolism proteins identified include: glucose 1,6 bisphosphate synthase, glycogen phosphorylase, and pyruvate kinase. The proteome of CLDs in enterocytes following a high fat dietary challenge highlights key proteins involved in the process of dietary fat absorption.
MED28 Regulates Cell Growth through HMG Box‐Containing Protein 1 (HBP1) in Human Breast Cancer Cells
Authors: Hsieh Nien‐Tsu , Huang Chun‐Yin , Chou Yu‐Hsuan , Ung Wong Ching , Lee Ming‐Fen
Published: June 2021

MED28 exhibits multiple cellular roles, including involved in transcriptional activation and as a merlin, Grb2, and cytoskeleton‐associated protein, which is highly expressed in several types of tumor. Recently, we have reported that MED28 regulates cellular migration and invasion in human breast cancer cells. Furthermore, overexpression of MED28 stimulates cell proliferation; however, the principal mechanism remains unclear. Therefore, the objective of this study is to understand the role of MED28 in the aspect of cell growth in breast cancer cells. MED28 suppression delays cell cycle progression in human breast cancer cells. MED28 appears to modulate cell growth through HMG box‐containing protein 1 (HBP1), a tumor suppressor which down‐regulates genes in G1 to S transition, and whose mutation or reduction is often manifested in invasive breast cancer. Overexpression of MED28 enhances cell cycle progression and downregulates HBP1 expression, whereas HBP1 cDNA partially blocks the stimulatory effect of MED28 on cell growth. Our data indicate that MED28 promotes cell cycle progression and stimulates cell proliferation, reinforcing the significance of MED28 in breast cancer development. (This work was supported by the grants, NSC 98–2320‐B‐309–002‐MY3 and NSC101–2320‐B‐309–001 to M‐F Lee, and CMU95–311 to C‐Y Huang.)
Identifying chemotaxis protein‐protein interactions in Epulopiscium sp. Type B using a yeast two hybrid system
Authors: Levesque Brié , Piefer Andrew J.
Published: June 2021

Epulopiscium sp. Type B (Epulo) is a large symbiotic aquatic bacterium that resides in the gut of surgeon fish. From genomic data, Epulo is predicted to have a chemotactic sensory and motility system similar to Escherichia coli and Bacillus subtilis. Our experiments are designed to test the protein‐protein interactions between methyl accepting chemotaxis proteins (MCPs) and the Che proteins, CheA (a sensory kinase) and CheW (a coupling protein), through the use of a yeast‐2‐hybrid (Y2H) genetic assay. We hypothesize based on genetic homology that Epulo chemotaxis proteins will behave in a similar fashion to their orthologs in B.subtilis and E.coli. The Y2H tests protein‐protein interactions through metabolic and drug selection in the yeast Saccharomyces cerevisiae. The genes in question; MCP3, MCP6 and CheW will be cloned into pGADT7 vector while CheA will be inserted into pGBKT7. Preliminary results confirmed an interaction between CheA and CheW; other interactions have yet to be detected; possibly because CheW is mildly toxic to the yeast strain Y187. Alternatively, a CheA+CheW+MCP ternary complex may be necessary for MCP binding.

This research is funded by a Hartwick College Faculty Research Grant awarded to AJP.
Osteogenic activity of milk thistle extract after ovariecotmy to dampen estrogen deficiency‐induced osteoporosis
Authors: Kim Jung‐Lye , Kim Yunho , Lee EunJung , Kang Young‐Hee
Published: June 2021

Bone integrity abnormality and imbalance between bone formation by osteoblasts and bone resorption by osteoclasts results in metabolic bone diseases such as osteoporosis. This study evaluated the suppressive effects of milk thistle extract (MTE) on bone loss. C57BL/6 female mice were ovariectomized (OVX) as a model for postmenopausal osteopenia and administered MTE (10 mg/kg per day) or MTE component silibinin orally for 8 weeks. The sham‐operated mice served as controls. MTE and silibinin enhanced alkaline phosphatase activity of osteoblasts but reduced tartrate resistant acid phosphatase (TRAP) activity of osteoclasts. The osteoprotective effects of MTE were comparable to those of estrogenic isoflavone. Low‐dose combination of MTE and isoflavone had a pharmacological synergy that may be useful for osteogenic activity. The treatment with nontoxic MTE and silibinin in ovariectomized mice improved femoral bone mineral density and serum receptor activator of nuclear factor‐κB ligand/osteoprotegerin ratio, an index of osteoclastogenic stimulus. In addition, MTE administration inhibited bone loss of the femur induced by ovariectomy and induction of cathepsin K necessary for osteoclast activitybone resorption. Collectively, oral doses of MTE in the preclinical setting are effective in preventing estrogen deficiency‐induced bone loss.
Food insecurity in Palestinian and Iraqi refugees in Lebanon
Authors: Ghattas Hala , Sahyoun Nadine R , Seyfert Karin , Sassine AnnieBelle J
Published: June 2021

The Middle East region has 1.8m refugees, the majority of which are Palestinians and Iraqis. In Lebanon, their legal status is compromised resulting in high rates of poverty and potential vulnerability to food insecurity. We aimed to assess food insecurity and its predictors in two surveys conducted in Palestinian (n=2575) and Iraqi (n=630) refugee households living in Lebanon. Food security was assessed using a 6‐item food security survey module. The prevalence of food insecurity was 41.2% in Palestinian and 35.5% in Iraqi households and that of severe food insecurity (SFI) was 20.8% and 45.9%, respectively. In multivariate logistic regression, the major variables associated with SFI in Palestinians and Iraqis were self‐reported mental health (OR=2.8, p=0.000; OR=1.7, p=0.019) respectively, and ownership of food‐related assets (OR=0.71, p=0.003; OR=0.75, p=0.039). Poverty, acute illnesses, and head of household primary education were associated with SFI in Palestinians, whereas number of children under‐15 and poor housing quality predicted SFI in Iraqis. Severely food insecure households in both refugee populations reported a significant reduction in intakes of fruits, meat and dairy products. Future interventions on food insecurity in Palestinian and Iraqi refugees should address mental health issues, education and dietary diversity.

This research was funded by the EU and UNHCR, Lebanon.
A prospective study of body image dissatisfaction and BMI change in school children
Authors: Duchin Ofra , Mora‐Plazas Mercedes , Marin Constanza , de Leon Carlos Mendes , Lee Joyce M , Baylin Ana , Villamor Eduardo
Published: June 2021

Objective

To examine the associations of body image dissatisfaction (BID) with body mass index (BMI) change in a longitudinal study of school children from Bogotá, Colombia.

Methods

We recruited 630 children 5–12 y‐old from public schools in Bogotá, Colombia and measured their height and weight annually for a median 2.5 y. At recruitment, BID was estimated from children's ratings of Stunkard‐style scales depicting progressively larger body figurines as current minus desired body image. Sex‐specific BMI‐for‐age curves were estimated by baseline BID levels, using mixed effects models with restricted cubic splines.

Results

In multivariable analyses, thin boys who desired to be thinner gained 4.4 kg/m2 more BMI from ages 6–14 y than boys without BID (P= 0.009). Overweight boys who desired to be larger or thinner gained significantly more BMI than boys with no BID. Thin girls who desired to be larger or thinner gained significantly less BMI than girls with no BID, but overweight girls who desired to be larger gained 4.7 kg/m2 more BMI than girls without BID (P=0.001). BID was not related to BMI change in normal‐weight children.

Conclusions

BID plays a sex‐specific role in BMI change among school children.

The Bogota School Children Cohort is currently sponsored by the ASISA Research Fund at the University of Michigan

Grant Funding Source: The Bogota School Children Cohort is currently sponsored by the ASISA Research Fund at the University of Michigan
When obesity begins: anthropometric and demographic characteristics of California WIC child participants
Authors: Papathakis Peggy Callaghan , Nazmi Aydin , Phelan Suzanne , Engle Patrice
Published: June 2021

Aim

to determine rates of overweight (OW) and obesity (OB) by demographic and anthropometric factors in children enrolled in 2004–2009 in San Luis Obispo and Santa Barbara county WIC programs.

Methods

longitudinal measurements were collected on 60,190 mother/child pairs. Using CDC growth standards, weight‐forlength z‐ scores (WLZ) were calculated from birth through 24 mo; BMI z‐ scores (BMIZ) were calculated from 24 to 57 mo. Proportion OW and OB were defined as WLZ and BMIZ ≥1.036 (>;85%tile)and >;1.645 respectively (>;95%tile).

Results

The overall prevalence of OW/OB was 28.9%/13.2% at 12 mo, 27.8%/12.4% at 24 mo, and 40.1%/22.2% at 57 months. At 12 mo, OW was significantly (p<0.05) higher in children who were male (30.6 vs 27.2%), non‐white Hispanic (29.5 vs 26.3% in whites) and formula fed for 6 months (39.0 vs 25.5% in breastfed). OW children had mothers who were <100% of poverty level (29.5 vs 27.1% in 185% poverty), with no education (33.8 vs 26.3% in 13+ yr education) and BMI >;30 (33.6 vs 24.4% in BMI 18.5–24.9). Similar demographic characteristics were observed at 57 months and for obese children.

Conclusion

Obesity begins in early life and is particularly prevalent among low income, non‐white Hispanic children whose mothers are obese and with low education.
Dietary patterns of Korean elementary school students in Incheon region
Authors: Kim YoungA , Cho Bong Hee
Published: June 2021

This study was conducted to provide basic information for enhancing Nutrition Education of Korean elementary school. 757 upper grade elementary school students were cross‐sectional surveyed in the region of Incheon, one of 5 major cities in Korea. A questionnaire was used for the survey, and the data was analyzed with SPSSWIN 18.0 program. The investigated students were divided in 3 groups according to Body Mass Idex(BMI). 3.5% were low weight(LW), 83.8% were normal weight(NW) and 12.7% were over weight(OW). In the aggregate, 82.3% of students ate 3 times a day, while 5.4% of students ate more than 4 times, 11.9% ate twice. The rate that students ate breakfast every day were overall 69.4%, while it grew down as their grade got higher (p<.001). And the rate was found 66.2% when their mother has a job, while it was 74.3% as their mother were housewives (p<.05). The ratio to have lunch every day was 88% and it was higher than other meals’ cases, by virtue of school meal systems. 62.9% of students ate more amount of meal at dinner than breakfast or lunch. And it was found that students with higher BMI tended to have fewer amount of dinner since the rate was significantly different by BMI as 84% for LW, 64% for NW, 49.4% for OW (p<.01). As for the snacks, 89.8% of students had snacks once or twice a day, and even 6.1% of students had snacks more than 3 times a day. The most frequent time slot for snacks was between lunch and dinner (66.6%). The girls’ most favorite snacks were fruits or fruit juice, while the boys’ most favorite ones were breads (p<.001). It was found that 44.1% of students had unbalanced eating habit, and the most dislikable foods were beans(40.8%), fishes(22.9%), vegetables(19.2%). There was a significant difference about unbalanced eating habit according to BMI (56.0%, 46.1%, 26.4% in LW, NW, OW, respectively) (p<.001). Therefore, nutrition education programs should be designed considering students’ characteristics such as grade, gender, BMI, etc.
Permethrin, a pyrethroid insecticide, potentiates adipogenesis in 3T3‐L1 adipocytes
Authors: Kim Jonggun , Park Yooheon , Yoon Kyung Sup , Clark John M , Park Yeonhwa
Published: June 2021

Permethrin belongs to a family of pyrethroids, naturally occurring insecticides, originating from Chrisanthemum flower heads. It is believed that pyrethroid insecticides have a relatively low toxicity due to limited absorption and fast metabolism, however, the potential link between prolonged exposure to environmental chemicals including pyrethroid insecticides and increased incidence of obesity has been suggested. Thus, the purpose of this study is to determine the effects of permethrin on adipogenesis using 3T3‐L1 adipocyte model. 3T3‐L1 cells were differentiated into adipocytes with 0, 10, and 20 μM permethrin for 6 days. Permethrin treatment at 10 and 20 μM dramatically increased triglyceride deposition compared to control. Expressions of major adipogenic genes, CAATT element binding protein‐α, peroxisome proliferator‐activated receptor‐γ, and fatty acid synthase, and expression of adipose triglyceride lipase were increased by permethrin. However, leptin mRNA expression was not changed by permethrin treatments. These data indicate permethrin could attribute to increased adiposity and potentially impair leptin signaling.
Higher protein preterm formula increases weight gain
Authors: Simpson Shelby , Kennedy Tay , Larzelere Robert
Published: June 2021

This study examined weight gain and growth in head circumference in 95 low birth weight infants (range 609–2495 grams) in a regional NICU before (n= 51) and after transition to a higher protein formula (n = 44). Multilevel modeling was used to predict the trajectory of gains in weight and head circumference based on the number of days the infants received traditional preterm formula, high protein formula or breast milk and/or a preterm discharge formula. Controlling for baseline growth (breast/discharge group) and predictors of growth trajectory including birth weight, gender and necrotizing enterocolitis, the high‐protein formula predicted an additional increase in weight (11 extra grams per day, t (897) = 6.38, p < .001) and a marginal additional increase in head circumference (.017 cm, t [1177] = 1.87, p = .06). In contrast, the traditional formula predicted an additional decrease in weight (−6.6 fewer grams per day, t [897] = −6.84, p < .001), but an additional increase in head circumference (.014 cm, t [1177] = 2.54, p < .05). Thus premature infants gained more weight from the high‐protein formula than from the traditional formula under conditions that likely had similar selection biases, but were equivalent in increasing head circumference.
Barley beta‐Glucan Consumption Decreases Serum Cholesterol Level and Increases 7 alpha‐Hydroxy‐4‐Cholesten‐3‐One Level in Hypercholesterolemic Adults
Authors: Wang Yanan , Harding Scott E. , Tosh Susan M. , Jones Peter J. , Ames Nancy P.
Published: June 2021

Interrupting bile acid metabolism has been proposed as a possible mechanism of cholesterol‐lowering effects of barley β‐glucan in humans. Measuring bile acid intermediate 7α‐hydroxy‐4‐ cholesten‐3‐one (7αHC) provides an indication of bile acid synthesis rate. The aim of this study was to determine whether interruption of bile acid metabolism is responsible for the cholesterol‐lowering effect of barley β‐glucan. In a controlled, four phase crossover trial, mildly hypercholesterolemic subjects (n=29) were randomly assigned to receive breakfast containing 5g low molecular weight (MW), 3g high MW, 3g low MW barley β‐glucan or a control, each for 5 weeks. Serum total cholesterol levels were reduced 0.39mmol/l, 0.55 mmol/l and 0.48mmol/l and LDL levels were reduced 0.20mmol/l, 0.31 mmol/l and 0.25mmol/l from baseline by these three barley treatment diets, respectively (p<0.05). 3g high MW resulted in greatest reduction in serum cholesterol and LDL levels. 7αHC levels were increased by 18.3 %, 39.7% and 32.2% by these three barley treatments, respectively, compared with control (p<0.05). For between‐group differences, 3g high MW increased 7α HC significantly compared to control (44.1 mmol/l vs.39.0 mmol/l). In conclusion, high MW barley β‐glucan resulted in greater reduction in cholesterol levels and increased 7α HC levels compared to low MW. Interference of bile acid metabolism may be the mechanism of cholesterol lowering for barley β‐glucan. Increasing MW may increase the cholesterol‐lowering bioactivity of barley β‐glucan. Supported by Growing Forward Agriculture and Agri‐Food Canada
Fermented barley averts diet induced obesity via modulating the lipid metabolic gene expression in Sprague dawley rats.
Authors: Pichiah P.B. Tirupathi , Moon Hye‐Jung , Moon Yeon‐Jeong , Cha Youn‐Soo
Published: June 2021

This study investigated the anti‐obesity effect of fermented barley on diet induced obesity in Sprague dawley (SD) rats. Animals were randomly divided into 6 groups (n=6); ND‐C, ND‐SH, normal diet plus guar gum or 1500 mg/kg bodyweight (BW) fermented barley group respectively; HD‐C, high fat diet plus guar gum; HD‐SL, HD‐SM and HD‐SH, high fat diet plus 500, 1000 and 1500 mg/kg BW fermented barley respectively. Guar gum supplementation was equivalent to the amount of soluble fiber in the fermented barley. After 19 weeks animals were sacrificed, serum lipid profile, leptin, ghrelin and hepatic lipid metabolic gene expression level were examined. A significant reduction in bodyweight, bodyweight gain, BMI, obesity index, epididymal adipose tissue weight, feed and energy intake was observed in HD‐SH group compared to HD‐C and HD‐SL group. Serum leptin decreased significantly in a dose dependent manner. Serum triglyceride and ghrelin level did not show significant change compared to the control group. Total cholesterol and LDL‐cholesterol tended to decrease; HDL‐cholesterol did not show significant changes. The mRNA expression of FAS was significantly decreased; ACO, PPARγ and PPARα increased dose dependently. CPT1α level was at similar level in all fermented barley group. These data indicated that fermented barley exerts an anti‐obesity effect against diet induced obesity conditions in rats.
Suppression of TPA‐induced Invasion of HepG2 cells by Glyceollins
Authors: Kim Jong‐Sang , Park In Sil , Kim Hyo Jung , Kim Sung Soon , Kang Hye Ryung , Kim Bo Ram , Zakaroba Alexandra , Seo Ji Yeon
Published: June 2021

Our previous study demonstrated that glyceollins (mixture of isomers I, II, and III), prenylated isoflavones derived from daizein in soybean elicited by fungi, had cancer preventive effect and antitumor activity. However, it is not known whether glyceollins inhibit the invasion of cancer cells. The purpose of this study was to assess the effect of glyceollins on the invasion of human hepatoma cells (HepG2) by 12‐O‐tetradecanoylphorbol 13‐acetate (TPA). Effects of glyceollins on cancer cell invasion were evaluated using wound healing and transwell assays. The effect of glyceollins on the expression of matrix metalloproteinase (MMP)‐ 9, mitogen‐activated protein kinases (MAPKs), AKT, and nuclear factor‐kappa B (NF‐kappa B), was investigated with gelatin zymography and western blotting. Glyceollins significantly suppressed TPA‐induced cell invasion in a dose dependent manner, which was accompanied with suppression of TPA‐induced MMP‐9 expression and activation of NF‐kappa B. Taken together, our results suggest that glyceollins inhibit the metastasis of human hepatoma cells by suppressing the expression of MMP‐9 which is mediated by NF‐kappa B signaling pathway.
Impact of high saturated lipids diet on serum fatty acid profile in rats.
Authors: Perris Paula Daniela , Silva Carolina , Fernandez Inés , Mambrin Cecilia , Slobodianik Nora , Feliu María Susana
Published: June 2021

Objetive

to examine the effect of high saturated lipids diet on serum fatty acid profile. Weaning Wistar rats were fed with experimental diet containing 45kcal% fat provided by butter, complete in all nutrients. Group S1 received the diet for 10 days; S2 for 40 days; S3 received stock diet for 30 days and then 40 days of experimental diet. Control groups (C1, C2, C3) received normocaloric diet for the same time. Serum fatty acid profile was determined by gas chromatography. Results (area%±SD) were:

S 1
C1
S 2
C 2
S 3
C 3

Palmitic
18.10 ±6.48
17.84 ±2.12
20.63 ±2.35 *
16.08 ±2.15
17.25 ±2.15 *
16.44 ±1.62

Oleic
19.12 ±5.18 *
10.60 ±2.01
20.37 ±2.06 *
11.29 ±2.27
17.71 ±2.85 *
10.08 ±1.34

Linoleic
8.91± 1.79*
19.99 ±4.68
9.79± 1.04*
17.45 ±4.11
8.38± 0.79*
17.82 ±3.38

αLinolenic
0.49± 0.32*
1.02± 0.46
0.44± 0.13*
0.81± 0.22
0.43± 0.11*
0.93± 0.18

Arachidonic
6.36± 1.45
8.59± 2.15
10.77 ±1.05 *
16.32 ±4.27
17.52 ±2.30 *
22.78 ±1.98

EPA
0.93± 0.82
0.83± 0.44
1.01± 0.15
1.13± 0.48
2.10± 0.20
1.83± 1.17

DHA
1.22± 0.30
1.25± 0.24
2.76± 0.35
2.22± 0.75
1.76± 0.78
2.17± 1.10

p<0.01

Experimental groups showed higher serum oleic acid levels with lower α‐linolenic and linoleic acids levels compared with control groups. Serum fatty acid levels changed in response to high fat% of experimental diet, the fatty acid profile, ‐which promotes ω9 pathway and decreases essential fatty acids‐, and to the time of administration.

Supported by UBACyT 20020100200078
TOLL‐LIKE RECEPTOR 4 (TLR4) MEDIATES ENDOTHELIAL DYSFUNCTION DURING TYPE I DIABETES
Authors: Carrillo‐Sepulveda Maria Alicia , Nunes Kenia Pedrosa , Webb R. Clinton
Published: June 2021

Endothelium dysfunction is one of the key events in diabetes, leading to progression of cardiovascular complications; however the underlying mechanism still remains elusive. TLR4 is a critical mediator of innate immunity, is expressed in vascular cells and has been reported to be activated during diabetes. We hypothesized that TLR4 signaling is activated in endothelial cells (EC) under hyperglycemic condition which causes endothelial dysfunction. TLR4 expression was robustly increased in EC primary cultures stimulated with 25mM high glucose for 48h [5.23±0.64 vs 2.95±0.37 normal glucose (NG), p<0.05] as well as MyD88, a downstream adapter protein of TLR4 signaling, (3.78±0.36 vs 1.44±0.55 NG, p<0.05). To test whether TLR4 contributes to endothelial dysfunction, streptozotocin (STZ)‐induced diabetic rats (65mg/kg, 5 weeks) were treated with a TLR4 signaling inhibitor (CLI‐095, 50ug/day) in the last 14 days. Aortas were isolated for functional studies and concentration‐responses to acetylcholine (Ach) were performed. Impaired Ach‐induced vascular relaxation observed in vessels from STZ rats compared to control (%47.17±1.74 vs. 75.05±4.34), p<0.05) was prevented with CLI‐095 treatment (%82.93±3.3, p<0.05). These results suggest that hyperglycemia upregulates endothelial TLR4 signaling which in turn contributes to endothelial dysfunction.
Non‐canonical MMP1‐PAR1 signaling drives vascular smooth muscle cell de‐differentiation and restenosis
Authors: Austin Karyn , Javid Golrokh , Kuliopulos Athan
Published: June 2021

Cardiovascular disease remains the leading cause of death in the United States. Aberrant vascular repair following plaque rupture or iatrogenic intervention is mediated by smooth muscle cells (SMCs). SMC proliferation, migration, and de‐differentiation from a contractile to a synthetic phenotype are crucial contributors to the pathology associated with maladaptive remodeling. The GPCR, protease activated receptor‐1 (PAR1), has been implicated in both vascular remodeling and atherothrombosis. Despite the well documented contribution of PAR1 to arterial thrombosis, it is unclear whether PAR1 promotes or mitigates the hyperplastic response to arterial injury. We sought to determine whether inhibition of upstream activating proteases of PAR1, matrix metalloprotease‐1 (MMP‐1) and thrombin, would have differing effects on SMC behavior and restenosis. We describe a previously unknown pathway through which MMP‐1 drives intimal hyperplasia and leads to SMC de‐differentiation through biased agonism of PAR1. In contrast, thrombin‐PAR1 signaling promotes a contractile phenotype. Also, pharmacologic blockade of MMP‐1 suppresses intimal hyperplasia following arterial injury. These data suggest a new strategy for targeted PAR1 inhibition that does not rely on receptor antagonism, but instead directly targets the detrimental activating protease. NIH grants F30 HL108590 (to K.M.A.), R01 HL64701 (to A.K.)
Selective Estrogen Receptor Modulators Resveratrol, Genistein, Tamoxifen and Pterostilbene down‐regulate the Expression of Estrogen‐Regulated Trefoil Factor 1 (TFF1) Gene in MCF‐7 Breast Cancer Cells
Authors: Bhatt Vrushank Dharmesh , Ratna Warren N
Published: June 2021

The human trefoil factor 1 (TFF1) is a small cysteine‐rich secretory protein expressed in the breast tumor in response to estrogen. The selective estrogen receptor modulators (SERMs) such as tamoxifen and raloxifene provide an effective treatment for estrogen receptor (ER) positive breast cancers. Our studies indicate that these SERMs at pharmacological concentrations modulate the expression of TFF1. The MCF‐7 breast cancer cells were treated with increasing concentrations of estrogen and SERMs such as resveratrol, genistein, tamoxifen and pterostilbene, and its effects were studied using RNA and protein analyses. The cell viability studies using estrogen showed increased cell growth even at low concentrations whereas the SERMs showed a concentration dependent cell growth arrest and death by apoptosis. Reverse Transcription‐Polymerase Chain Reaction (RT‐PCR) and Northern blot analyses indicated decreased levels of transcription of TFF1 mRNA. Western blot analysis showed a corresponding decrease in the expression of the protein. The above results indicate the potential use of these SERMs for the down‐regulation of TFF1 expression and consequently the regression of ER positive breast tumors. This work was supported by science research funds from Arnold and Marie Schwartz College of Pharmacy, Long Island University, NY.
Rho kinase (ROCK) inhibition counteracts tyrosine kinase inhibitor‐induced rise in arterial pressure and renal vascular resistance
Authors: Grisk Olaf , Meissner Thomas , Donner Alexander , Braun Diana , Zimmermann Uwe , Rettig Rainer
Published: June 2021

The VEGF antagonistic tyrosine kinase inhibitor sunitinib is used to treat renal cell carcinoma but causes severe arterial hypertension limiting its therapeutic utility. We investigated the in vitro and in vivo effects of sunitinib on renal vascular function and tested if ROCK inhibition prevents sunitinib‐induced hypertension in rats. VEGF dilated isolated rat renal resistance vessels (log EC50 −11.1 ± 0.25 mol/l). In in vitro experiments, sunitinib did not affect basal vascular tone and endothelium‐dependent vasodilation but significantly reduced agonist‐induced vasoconstriction in isolated renal resistance arteries. Chronic sunitinib treatment (15 mg*kg−1*d−1) increased arterial pressure by 25 mmHg and renal vascular resistance (RVR) by 4 mmHg*ml−1*min*g−1 within 4 days (p < 0.05). Sunitinib‐treated rats showed blunted endothelium‐dependent renal vasodilation and reduced angiotensin II‐induced renal vasoconstriction while α1‐adrenoceptor‐dependent vasoconstriction was unaffected. ROCK inhibitor treatment prevented the sunitinib‐induced rises in arterial pressure and RVR but not renal endothelial dysfunction. ROCK‐inhibition may be useful to mitigate tyrosine kinase inhibitor‐induced hypertension thereby improving the treatment of renal cell carcinoma.
MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) DECREASES NEUROINFLAMMATION IN THE SOLITARY TRACT NUCLEUS (NTS) OF SPONTANEOUSLY HYPERTENSIVE RATS (SHR).
Authors: Freiria‐Oliveira Andre H , Blanch Graziela T , Li Hongwei , Colombari Debora S A , Colombari Eduardo , Sumners Colin
Published: June 2021

Increasing MIF levels in the NTS reduces blood pressure in SHR. Since inflammation has an important role in hypertension we investigated whether increasing MIF in the NTS of SHR alters inflammatory cytokine levels and microglial activation. Male SHR and normotensive (NT) rats (8 weeks old) were microinjected in the NTS with either AAV2‐CBA‐MIF or AAV2‐CBA‐eGFP to elicit chronic increases in MIF and GFP (control) expression. Thirty days later, we analyzed inflammatory cytokine mRNAs or microglial activation in the NTS. Baseline levels of pro‐inflammatory cytokine (PIC) mRNAs such as IL‐1β, IL‐6 and TNFα were significantly greater (0.5 to 3 fold) in the NTS of SHR vs. NT rats (p<0.05). The baseline level of IL‐10 mRNA (anti‐inflammatory cytokine) was significantly reduced by 3 fold in SHR vs. NT rats. MIF over expression in the NTS did not alter cytokine mRNA levels in NT rats, but in SHR normalized the levels of PIC and IL‐10 mRNAs in the NTS towards those observed in NT rats (p<0.05). Qualitative assessment of microglial activation via Iba‐1 immunohistochemistry indicated greater microglial activation in the NTS of SHR vs. NT rats, and this was reversed by MIF over expression. The results suggest that MIF over expression in the NTS of SHR can reduce neuroinflammation at this site, and this may contribute to the reduce blood pressure observed in these animals.

Grant Support: FAPESP/PRONEX, CNPq, NIH (HL076803)
Tachycardia promotes Mitochondrial DNA Damage by JNK Translocation to Mitochondria
Authors: Jen Nelson , Li Rongsong , Hsiai Tzung
Published: June 2021

Atrial fibrillation (AF) is the most common cardiac arrhythmia that affects 5% of the population over the age of 65. The rapid and irregular pulse rates associated with AF have been reported to lead to altered hemodynamics by irregular stroke volumes and turbulent flow. Shear stress, the viscous drag force of blood flow, regulates endothelial function and is intimately related to vascular oxidative stress and inflammatory responses. We hypothesize that the irregular ventricular response associated with AF contributes to pro‐inflammatory states that lead to mitochondrial dysfunction in endothelial cells. A dynamic flow system was used to implement pulsatile flow in response to tachycardia at arterial carotid bifurcations. Human aortic endothelial cells (HAEC) were exposed pulsatile shear stress at 23±5 dyne·cm−2 at 140 beats per minute (PSS140, CL=429). PSS140 induced JNK activation at the mitochondria at 30 minutes (1.85‐fold increased compared to static, p<0.05, n=5) which as accompanied by an increased in MitoSOX Red intensities. PSS140 also increased mitochondrial DNA damage (as measured by 4,977‐bp deletion) and autophagy. Inhibition of JNK and antioxidant treatment attenuated these responses. When taken together, these findings suggest that JNK activation mediates rapid‐pulse rate induced cellular redox states which
The association of plasma homocysteine and cysteine with antioxidant enzyme activities in patients with colorectal polyps and colorectal cancer
Authors: Chen Fang‐Pei , Huang Yi‐Chia , Lin Chun‐Che , Wang Hwei‐Ming , Chiang Feng‐Fan
Published: June 2021

Colorectal polyps are considered to be precursors of colorectal cancer, especially of adenomatous polyps. Cysteine is synthesized by the transsulfuration pathway of homocysteine, and further contributed to glutathione. Although increased homocysteine has been shown to be associated with the development of colorectal polyps and carcinogenesis, the role of cysteine is not cleared. The purpose of this study was to investigate the association of plasma homocysteine and cysteine with glutathione‐dependent antioxidant enzymes [i.e., glutathione peroxidase (GPx) and glutathione S‐transferase (GST)] in patients with colorectal polyps or colorectal cancer. 70 subjects with adenomatous polyps, 40 subjects with hyperplastic polyps and 168 patients with colorectal cancer were recruited. Fasting plasma homocysteine and cysteine concentrations in patients with colorectal cancer were significantly higher than subjects with adenomatous and hyperplastic polyps. There was no significant association of homocysteine with GPx and GST activities in all subjects. However, cysteine significantly correlated with GPx and GST after adjusting for age, gender, serum creatinine and smoking in subjects with adenomatous and hyperplastic polyps. Higher cysteine rather than homocysteine is an important contributing factor in the elevation of GPx and GST activities in subjects with colorectal polyps.
MYLOID DIFFERENTIATION FACTOR 88 (MYD88) DOES NOT CONTRIBUTE TO HYPERTENSION OR ENDOTHELIAL DYSFUNCTION PRODUCED BY ANGIOTENSIN II
Authors: Gomolak Jessica R , Faulkner Jessica L , Didion Sean P
Published: June 2021

Although a role for adaptive immunity has begun to emerge in hypertension, much less is known regarding the potential role of innate immunity in hypertension. The goal of this study was to test the hypothesis that deficiency of MyD88 (a key mediator of toll‐like receptor signaling) limits the development of hypertension and endothelial dysfunction produced by Angiotensin II (Ang II). Blood pressure (BP) and vascular responses were examined in wild‐type (WT) and MyD88‐deficient (MyD88−/−) mice infused with Ang II (1000 ng/kg/min via osmotic minipump for 14 Days). Ang II produced a similar degree (P>;0.05) of hypertension and cardiac hypertrophy in WT and MyD88−/− mice. Ang II infusion was associated with endothelial dysfunction in carotid arteries from Ang II‐infused WT and MyD88−/− mice as compared to their respective controls (p<0.01). Ang II produced a similar degree of endothelial dysfunction in carotid arteries from WT and MyD88−/− mice. Contractile responses to U46619 were similar in all groups. Contrary to our hypothesis, these data suggest that MyD88, a major component of innate immune signaling, does not contribute to the hypertension or endothelial dysfunction produced by Ang II. Supported by NIH HL‐089884 & HL‐107632.
Accelerators and brakes in CO2 chemosensitive neurons
Authors: Quintero Maria Carolina , Cordovez Juan Manuel , Putnam Robert W.
Published: June 2021

The cellular factors that determine the magnitude of the firing rate response of chemosensitive neurons to increased CO2/H+are typically attributed to acid‐induced increased firing rate pathways (accelerators). However, a marked reduction in the CO2/H+‐induced increased firing rate during postnatal development has been observed in neurons from the locus coeruleus (LC). This response has been hypothesized to be due to the development of a decelerating pathway that arises from CO2/H+ activation of Ca2+ channels, which in turn induces the Ca2+‐activation of KCa channels. In order to investigate the contribution of such a braking pathway in the magnitude of the firing rate in individual neurons, we have developed a preliminary computational model of excitable single neuron that simulates the voltage‐gated currents as well as the pH and Ca2+ sensitive K+ currents using the Hodgkin‐Huxley formulation. In our simulations we found that inhibition of the L‐type Ca2+current led to a larger hypercapnic‐induced firing rate response. These findings constitutes theoretical evidence that the cellular factors limiting the firing rate response of chemosensitive neurons are associated with acid activated Ca2+ and Ca2+ activated KCa currents, allowing us to conclude that a braking pathway may play a more significant role in setting neuronal chemosensitivity. Supported by NIH Grant HL56683.
The differential accumulation of doxorubicin in the soleus and gastrocnemius muscle of the rat
Authors: Fabris Sergio Antonio , Chenard Pamela , Anderson Lindsay , MacLean David
Published: June 2021

The purpose of the present study was to examine intramuscular (IM) doxorubicin (DOX) levels in the soleus (S) and gastrocnemius (G) muscles of Sprague‐Dawley rats following the administration of DOX (1.5 or 4.5 mg/kg). Muscles were collected at 24, 48, 72 and 192 hrs post injection. DOX concentrations were elevated (P<0.05) as compared to control at all time points. Following the 1.5 mg/kg dose, concentrations peaked in both muscles after 48 hrs, but were higher (P<0.05) in the S (1.47±0.46 nmol/g) as compared to the G (0.78±0.24 nmol/g). Similarly, DOX was higher (P<0.05) in the S (0.40±0.13 nmol/g) than the G (0.08±0.02 nmol/g) after 192 hours. The 4.5 mg/kg dose resulted in similar peak DOX concentrations in both the G (2.43±0.52 nmol/g) and the S (2.46±1.35 nmol/g), but this occurred sooner (48 hr) in the S as compared to the G (72 hrs). DOX levels were not different between muscle groups following 192 hrs. These data not only support the notion that DOX accumulates in skeletal muscle in a dose dependent manor, but clearly demonstrate that fiber type differences are a factor in the rate of accumulation and removal or degradation of intracellular DOX. These findings have clinical implications as skeletal muscle may be an important compartment for the sequestering of DOX from the circulation, effectively limiting its therapeutic impact. Supported by NSERC.
Dissociation of recovery of muscle activation and force following a sustained maximal isometric contraction
Authors: Houle Katherine E , Kantner Bryan J , Brechue William F
Published: June 2021

To investigate time‐dependency of nervous system recovery following muscular contractions subjects (n=10) performed two bouts (B1, B2) of sustained maximal isometric handgrip contractions separated by 10‐min recovery. Force and surface EMG were collected continuously throughout contraction bouts but were analyzed at 15 sec intervals (1 sec segments). iEMG and mean power frequency (MPF) were calculated for the brachioradialis (BR), flexor carpi radialis (FCR) flexor carpi ulnaris (FCU), and flexor digitorum profundus (FDP) muscles. Muscle activation (iEMG‐MPF: BR:68–20%; FCR:72–16%; FCU: 65–20%; FDP:48–50%, respectively) and isometric force (88%) decreased following B1(120‐sec contraction). Following recovery, initial force of B2 was significantly less than B1 (23%); however muscle activation (iEMG) was similar to B1 for BR, FCR and FDP. Initial B2 FCU iEMG activity was lower but increased to B1 levels by 15 sec; remaining similar throughout. MPF was similar and decreased similarly over time in both bouts but tended to be higher at initiation of B2. The mechanical response was similar in both bouts despite differences in initial force generation (B1: y = −0.172x+22.7; R2=0.98; B2: y = −0.182x+17.9 R2=0.97). Incomplete recovery of force observed in B2 suggests interference in excitation‐contraction coupling while fatigue within each bout appears specifically related to changes in muscle activation.
MC3R activation does not attenuate pro‐inflammatory macrophage‐induced reductions in myoblast proliferation
Authors: Kump David S , Brooks Sarah , Kump Rachel A
Published: June 2021

Background

Activation of the melanocortin‐3‐receptor (MC3R) has potent anti‐inflammatory effect on macrophages. Acute muscle injury (AMI) is followed by an invasion of pro‐inflammatory macrophages (MΦ1).

Objective

The objective of this experiment was to determine whether 1) MΦ1would affect myoblast proliferation, a key step in recovering from AMI and 2) whether MC3R activation would alleviate the effects of MΦ1 on myoblast proliferation.

Methods and Results

RAW 264.7 MΦ (2.0 × 106 in a 10‐cm dish) were treated with 10 ng/ml for 24 hours. Conditioned media was added to 0.5 × 106 C2C12 myoblasts in a 24‐well plate for 48 hours, then proliferation of the myoblasts was measured via the MTT assay. Proliferation of myoblasts treated with MΦ1‐conditioned media was significantly lower than controls from non‐activated MΦ1. Alternatively, the MΦ1were scraped, and 5,000 MΦ1were co‐cultured with 0.5 × 106 C2C12 myoblasts in a 24‐well plate for 48 hours. Myoblasts in the presence of MΦ1also had significantly decreased proliferation, providing additional evidence that MΦ1 reduce myoblast proliferation. Pre‐treatment of the MΦ for 24 hours with 100 nM MT‐II, a MC3R agonist, did not attenuate the decreased proliferation. Current experiments are examining the possibility of a dose‐response relationship.

Supported by Winston‐Salem State University Department of Life Sciences.
The myosin converter region influences cross‐bridge stiffness and muscle kinetics
Authors: Swank Douglas , Glasheen Bernadette , Ramanath Seemanti , Wang Qian , Sheppard Debra , Riley Lauren
Published: June 2021

The converter has been proposed to be the myosin elastic element that sets cross‐bridge stiffness. According to most cross‐bridge models, the elastic element's stiffness should directly influence muscle fiber type force and power generation levels. We tested these hypotheses by using the natural variation in converter regions found in Drosophila muscle types. In Drosophila, myosin isoforms are generated by alternative splicing from a single myosin heavy chain gene. The converter region is encoded by five alternative exon versions. We expressed four non‐native versions in Drosophila indirect flight muscle (IFM) and found that two of the three versions, 11b and 11e (found in larval and adult body wall muscles) decreased IFM rigor stiffness by 63% and 43%, respectively. This supports the hypothesis that the converter is the critical elastic element of myosin. The converters also influenced IFM fiber and myosin kinetics. 11b, 11d, and 11e decreased the frequency at which maximum power was generated (fmax) by 28%, 40%, and 53%, respectively. All three decreased myosin attachment kinetics, however, 11d and 11e also increased detachment kinetics. Differences in force generation are minimal, likely due to myosin stiffness changes being offset by changes in myosin duty ratio. With little to no force differences, changes in muscle power almost directly correlated with fmax. All three lines displayed reduced wing beat frequency, 11 to 15% less than control lines, with flight performance better correlating with fiber fmax than cross‐bridge stiffness. Thus, the converter is critical for muscle fiber type diversity as it helps set myosin stiffness, cross‐bridge kinetics and muscle power generation. Supported by NIH grant R01 AR055611 to D.S.