bluuebunny/crossref_metadata_2025 huggingface dataset:
Maternal Serum Fatty Acids are Positively Associated with Systolic Blood Pressure Changes during Pregnancy: Longitudinal Study.
Authors: Farias Dayana , Rebelo Fernanda , Eshriqui Ilana , Lepsch Jaqueline , Pinto Thatiana , Franco‐Sena Ana , Lima Natália , Kac Gilberto
Published: June 2021

Aim

To evaluate if maternal serum Fatty Acids (FAs) are associated with Systolic (SBP) and Diastolic (DBP) blood pressure changes during pregnancy.

Methods

Prospective cohort of 225 healthy pregnant women followed at all pregnancy trimesters: 5th‐13th, 20th‐26th and30th‐36th weeks. SBP and DBP (mmHg) were measured twice at each follow‐up visit using an automatic sphygmomanometer (Omron HEM‐742 INT). The mean values at each trimester were used. Blood samples were collected after 12 hours of fasting. Serum FAs (µg/ml) were evaluated by a high‐throughput robotic direct methylation coupled with fast gas‐liquid chromatography. Longitudinal linear mixed‐effects models were adjusted for gestational age, socio‐economic, demographic, lifestyle, biochemical and anthropometric variables.

Results

We observed a positive and significant association of total saturated (β=0.004; 95% CI: 0.001‐0.008), monounsaturated (β=0.004; 95% CI: 0.0003‐0.009) and polyunsaturated n‐6 (β=0.005; 95% CI: 0.001‐0.009) FAs with SBP levels during pregnancy. The following fractions were associated with SBP: 16:0 (β=0.005), 18:0 (β=0.040), 20:0 (β=0.746), 16:1 n‐7 (β=0.028), 18:1 n‐7 (β=0.047), 18:1 n‐9 (β=0.006), 18:2 n‐6 (β=0.005), 20:3 n‐6 (β=0.046) (all p‐values<0.05). We did not observe significant associations between FAs and DBP and between polyunsaturated n‐3 FAs and BP changes during pregnancy.

Conclusion

Maternal serum concentrations of saturated, monounsaturated and polyunsaturated n‐6 FAs were positively associated with SBP changes during pregnancy.

Support: CNPq and FAPERJ
The effects of consuming high protein solid vs. beverage breakfasts on changes in body weight and body composition in overweight, breakfast skipping adolescents
Authors: Gwin Jess , Leidy Heather
Published: June 2021

Background

The consumption of high protein (HP) solid breakfast meals lead to improved appetite control and satiety; however, HP beverages elicit a blunted satiety effect. Purpose: To examine the long term effects of consuming HP Solid vs. Beverage breakfasts on body weight management in overweight breakfast skipping teens. Methods: Twelve overweight adolescents (age: 14 ± 1y; BMI: 93.1 ± 4 %ile; breakfast skipping: 6 ± 1 skipping occasions/wk) consumed either a 300 kcal HP (28 g Protein/28 g Carbohydrates/8 g Fat) Solid breakfast or HP Beverage breakfast for 12 wks. Specifically, the HP solid meals consisted of commonly consumed egg and dairy‐based breakfast foods, whereas the HP beverage meals were egg and dairy‐based breakfast shakes. Body weight, body composition, and appetite were measured during baseline and post‐study. Results: No main effects of time, treatment, or interactions were detected for weight, fat mass, or lean mass. Although no main effect of time or group were detected for percent body fat, a group by time interaction tended to occur (p=0.09). Post‐hoc comparisons demonstrated that the HP Solid breakfast led to greater changes in percent body fat (‐3.23 ± 1.9%) compared to the HP Beverage breakfast (+0.63 ± 0.9%; p=0.09).

Conclusions

The consumption of a high protein solid breakfast led to improvements in weight management in overweight, breakfast skipping adolescents, whereas a high protein beverage breakfast did not. Funding: NCRR/NIH M01 RR023940
Effects of Roux‐en‐Y gastric bypass on sugar‐induced hypertriglyceridemia
Authors: Surowska A , De Giorgi S , Theytaz F , Lecoultre V , Campos V , Giusti V , Tappy L
Published: June 2021

Oral fructose increases postprandial blood triglyceride (TG) concentrations, possibly by stimulating intestinal de novo lipogenesis (iDNL) and decreasing extrasplanchnic TG clearance. We hypothesized that these effects of fructose may be abolished by Roux‐en‐Y‐gastric bypass (RYGB) due to bypass of the proximal small bowel. To evaluate this hypothesis, we studied 8 patients 12‐18 months after RYGB, and 8 age‐, weight‐ and gender‐matched controls (C). Each participant was studied on two occasions, over 6‐hours after ingestion of 1) a protein/ lipid meal (PL), and 2) a protein/ lipid/ glucose/ fructose meal (PLGF). Chylomicrons‐triglyceride secretion and clearance were assessed from postprandial incremental areas under the curve for TG and apoB48. Sugar‐related hyperlipemia was assessed from the difference between chylomicron‐TG iAUC after PLGF and after PL.

Postprandial chylomicrons‐TG iAUCs were, in RYGB: 16.8 ± 7.0 mmol/L*360min after PL vs 6.4 ± 5.7 mmol/L*360min after PLGF (NS), and in C: 28.4 ± 12.7 mmol/L*360min after PL vs 67.9 ± 23.0 mmol/L*360min after PLGF (p = 0.025). Sugar‐related hypertriglyceridemia was 2.1 ± 10.6 mmol/L*360min in RYGB vs 46.7 ± 15.0 mmol/L*360min in C (p = 0.03). ApoB48 peaked earlier in RYGB than in C, but post‐prandial iAUCs for apoB48 were not different in the two groups.

These results indicate that sugar induced hypertriglyceridemia was abolished after RYGB. A lower iDNL may possibly account for this effect of RYGB. An increased extrasplanchnic clearance of chylomicrons‐TG (as suggested by lower TG iAUCs together with unchanged apoB48 iAUCs) may also be involved.

This work was supported by the Swiss National Science Foundation
Relation between Candida spp in Dental Plaque and Nutritional Status in Kindergarten Children
Authors: Rodriguez Patricia , Mateo María , Iglesias María , Martinez Reinoso Josefina , Lifshitz Fima , Molgatini Susana , Argentieri Angela
Published: June 2021

Early childhood caries, an ecological disease, and obesity are related to dietary habits. We previously found an association among nutritional status (NS), prevalence of caries and microbiology composition of dental biofilm, that leads to a negative relationship between obesity and caries risk. Candida spp is a frequent commensal yeast of the oral microbiota; however, in the pediatric population is associated with presence of caries.

The aim of the present study was to determine the association between NS, Candida spp, and caries in preschool children. Fifty one children aged 3 to 6 years were were classified in adequate (A), overweight (OW) or obese (Ob) with Anthro software. Presence/absence of caries lesions, and Candida spp in the dental plaque were determined. Dental plaque samples were collected and seeded in selective medium CHROMagar Candida®.

Results

NS showed 49% of A, 29.4% OW and 21.6% Ob. Candida spp strains were isolated from dental plaque of (% and CI95%): 52 (32‐72) of A, 33 (12‐62) OW and 0(0‐28) Ob children (Fisher's test, p=0.0107), regardless the presence of caries. Althoug 36% of Ob children presented caries, we did not isolated Candida spp in dental plaque.

Conclusion

Colonization of dental plaque biofilm by Candida spp is related with nutritional status of preschoolers.

UBACyT 20720130100017BA.
Consuming Iron‐Biofortified Beans Reduces Time Spent in Discretionary Sedentary Activity in Iron‐Depleted Rwandese Women Compared to Conventional Beans
Authors: Luna Sarah , Lung'aho Mercy , Gahutu Jean Bosco , Haas Jere
Published: June 2021

Iron deficiency has been shown to negatively impact physical performance in laboratory‐based tests; however, little is known about its effects on free‐living physical activity. To test whether consuming iron‐biofortified beans reduces discretionary time spent in sedentary activity (MET=1), accelerometry data were collected in 111 Rwandese university women before and after a 20‐week iron‐biofortification feeding trial. From six‐day accelerometry data, we analyzed weekend values when participants had discretionary time. Metabolic equivalents (METs) were calculated with Crouter's refined 2‐regression model, and nonwear time was defined as vector magnitude = 0 for 60 consecutive minutes. Mixed effects models analyzing change in activity as the dependent variable were used to analyze the effect of consuming iron‐biofortified beans on time spent in different levels of physical activity. Women in both iron‐biofortified and control bean groups increased time spent in sedentary activity; however, those who consumed iron‐biofortified beans increased 102 min compared to 201 minutes in women who consumed the control bean (p=0.04). Change in hemoglobin from baseline to endline significantly predicted minutes spent in sedentary activity (estimate: ‐7.34 min per g/L Hb; p=0.04). The increase in discretionary sedentary activity between baseline and endline (both groups) may have been influenced by the nature and intensity of academic activities during the two periods; however, the effect of biofortification reducing time spent in sedentary activity is evident. Funded by HarvestPlus.
In Vitro Batch Fermentation Analysis of Wheat Dextrin and Partially Hydrolyzed Guar Gum ‐ Fermentation Kinetics and Prebiotics Effects
Authors: Carlson Justin , Slavin Joanne , Deng Ping , Swanson Kelly , Hospattankar Ashok
Published: June 2021

Scientific research demonstrates that two indigenous gut bacteria, Lactobacillus and Bifidobacterium can contribute to human health. The primary objective of this in vitro study was to quantitatively analyze at the genus level how wheat dextrin (WD) and partially hydrolyzed guar gum (PHGG) increased the levels of these two gut bacteria at 12 h and 24 h, via qPCR. Secondary objectives were changes in fecal pH, short chain fatty acids (SCFAs) and total gas volume produced. At 12 h WD was significantly more bifidogenic (9.50 CFU log10/mL) than PHGG (9.30 CFU log10/mL) (p<0.05), and also at 24 h WD (9.41 CFU log10/mL) compared with PHGG (9.27 CFU log10/mL) (p<0.05). WD produced less total SCFAs at both 12 h and 24 h (p<0.05), and produced significantly lower amounts of gas at 12 h and 24 h (p<0.05). Both PHGG and WD also promoted growth of Lactobacilli when measured at 12 h and 24 h compared with the 0 h analysis (p<0.05), indicating that both fibers are lactogenic. These results demonstrate prebiotic effect of WD and PHGG. Based on fermentation kinetics, PHGG is more rapidly fermented than WD, and both fibers show prebiotic effects as early as 12 h.

This study was supported by a research funding from Novartis Consumer Health Inc. NJ, USA.
Detection of Intimal Macrophages in Atherosclerotic Lesions Using a CD36‐Targeted Ligand Containing Nanoparticle
Authors: Dhanasekara Chathurika , Zhang Jia , Nie Shufang , Wang Shu
Published: June 2021

Objective

Atherosclerotic cardiovascular disease is the leading cause of death in the United States. Macrophages and lipid‐laden macrophages (foam cells) are the predominant immune cells in the lesions. We have synthesized biocompatible and biodegradable nanoparticles (NP), and we incorporated CD36‐targeting ligands on the surface of NP to make ligand‐NP (L‐NP). Our objective was to compare the target specificity of NP and L‐NP to macrophages in vitro and in LDL receptor null (LDL r‐/‐) mice.

Methods: THP‐1 derived macrophages were incubated with florescence‐dye labeled NP and L‐NP and target specificity was measured using an EVOS® Fluorescence Microscope. Six male 6 week old LDL r‐/‐ mice developed atherosclerotic lesions after feeding an atherogenic diet for 22 weeks. Fluorescence‐dye labeled NP or L‐NP were injected via tail veins. After 20 hours, fluorescence signals in aortic lesions were imaged in‐situ and in isolated aortas using an IVIS® Lumina XR imaging system. Then dissected aortas were embedded in O.C.T. compound and serially sectioned. Fluorescence signals in cross‐sections of aortas were taken using an EVOS® Fluorescence Microscope. Total Cy7 signal intensity in the cross‐sections of aortic lesions was analyzed using a NIH Image J software.

Results: L‐NP as compared to NP had higher binding affinity to THP‐1 derived macrophages

image

In‐vivo data showed that L‐NP had 1.4‐fold higher target specificity to aortic lesions than NP (p<0.001)

image

Conclusion

Our novel L‐NP has the potential for the prevention and treatment of atherosclerosis through targeting the macrophages in atherosclerotic lesions.

Grant Funding Source: NIH 1R15AT007013‐01
Alteration of Food intake, Inflammatory Response, and Insulin Resistance by Seaweeds with High‐Fat Diet in C57BL/6N Mice
Authors: Lee Yunkyoung , Oh Jihyun , Han Arum , Hwang Yeonji
Published: June 2021

Seaweeds contains various bioactive compounds and health promoting effects. Anti‐inflammatory and anti‐adipogenic effects of seaweeds have been intensively studied, but their mechanism(s) have not been clearly understood yet. In this study, we investigated that effects of four edible seaweeds (Undaria pinnatifida(UP), Laminaria japonica(LJ), Sargassum fulvellum(SF), Hizikia fusiforme(HF)) on alteration of food intake, obesity, insulin resistance, and inflammation by using in vivo and in vitro models. Six week‐old C57BL/6N male mice were fed with either low fat diet, high fat diet (HFD), or HFD+5% of each four edible seaweeds shown above for 16 weeks. Food intake/body weight and fasting blood glucose were measured every other day and every 4 weeks, respectively. Insulin tolerance test (ITT) was performed at 14 week. Additionally, bone‐marrow derived cells from 6 groups were isolated and differentiated to macrophage (BMDM) to evaluate the inflammatory response by LPS challenge. As a result, 5% seaweeds addition to HFD did not alter body weight changes compared to that of HFD fed group. Although food intake data suggested that there was a slight alteration of food intake in the beginning, the changes eventually disappeared. In spite of being obese, LJ contained HFD fed group demonstrated significantly improved insulin sensitivity than HFD only fed group based on ITT. LPS treated BMDM from LJ+HFD mouse expressed significantly less amount of IL‐6 compared to LPS treated BMDM from HFD group. In conclusion, LJ among 4 types of edible seaweeds demonstrated to have potential anti‐diabetic and anti‐inflammatory effects in diet induced obese model. This work was supported by NRF‐2013R1A1A1057573
Neuroprotective effects of hexane‐ and methylene chloride‐soluble fractions of Salicornia herbacea L. in mouse hippocampal HT22 cells
Authors: Kim Min Sun , Seo Ji Yeon , Kim Mi Hye , Woo Ji Eun , Kang Su Jin , Jo Gwang Rae , Ji Hyeon‐Kyeong , Erdenebileg Zolzaya , Jang Jin Ho , Kim Jae‐Sik , Suh Hwa Jin , Kim Jong‐Sang
Published: June 2021

Salicornia herbacea L. is one of the halophytic herbs that grow in salt marshes and on muddy seashores. It has been reported to exhibit their bioactivities including anti‐diabetic, antioxidant and anticancer activities. However, studies on the neuroprotective effect of this plant have not been conducted. Salicornia herbacea L. was extracted with 80 % ethanol, followed by further fractionation into five subfractions according to polarity; n‐hexane, methylene chloride, ethyl acetate, n‐butanol and water soluble fractions. The neuroprotective effect of plant extract and its solvent fractions was evaluated by their ability to counteract cell death and ROS generation induced by glutamate in murine hippocampal HT22 cell line. Our results indicated that n‐hexane and methylene chloride‐soluble fractions had the highest neuroprotective activities among 5 subfractions. These fractions also induced NAD(P)H:quinone oxidoreductase, an antioxidant marker enzyme, in a concentration‐dependent manner. These results warrant further mechanistic study and identification of active compounds in n‐hexane and methylene chloride‐soluble fractions of Salicornia herbacea L. (Supported by a grant (R0002736) from Regional Industry R&D Assistance Program (2013) of Ministry of Trade, Industry and Energy, Republic of Korea)
Shikonin Prevents Adipogenic Differentiation by Inhibiting Mir‐34a
Authors: Jang Young Jin , Ahn Jiyun , Jung Chang Hwa , Ha Tae Youl
Published: June 2021

Shikonin is a naturally occurring naphthoquinone pigment and a major component of Lithospermum erythrorhizon. Our previous study showed that shikonin inhibited adipocyte differentiation in 3T3‐L1 cells by suppression of ERK pathway. MicroRNAs (miRNAs) are key post transcriptional regulator of adipocyte differentiation. In present study, we investigated shikonin regulates adipocyte differentiation by targeting miRNA in 3T3‐L1 cells. 3T3‐L1 cells were differentiated by 3‐isobutyl‐1‐methylzanthine, dexamethasone, and insulin (MDI) for 8 days in the presence of 0‐2 μM shikonin. We found that mir‐34a was increased during adipocyte differentiation by MDI, and was suppressed by shikonin which are measured by quantitative real‐time PCR. mRNA expression of FKBP1B, one of target of mir‐34a suggested by bioinformatics studies was decreased during adipocyte differentiation and was recovered by shikonin which are reversely correlated with mir‐34a expression. Transfection of mir‐34a inhibitor suppressed MDI‐induced adipocyte differentiation which was analyzed by Oil Red O staining, and inhibited MDI‐induced PPARγ and C/EBPα expression. Suppression of mir‐34a also recovered MDI‐induced down‐regulation of FKBP1b mRNA expression. Increase of mir‐34a by transfection of mir‐34a mimic decreased FKBP1b mRNA expression in 3T3‐L1 preadipocytes. In conclusion, these results demonstrate that shikonin prevents adipogenic differentiation by blocking mir‐34a‐FKBP1B pathway, which could be a target for obesity prevention.
Effect of Long‐term Supplementation of Selected Medicinal Herbs on Activity of Intestinal α‐Glucosidase and Expression of SI‐complex in Rat Model
Authors: Lee Jung‐Yun , Kang Yu‐Ri , Choi Hwang‐Yong , Park Seung‐Hwan , Park Eun‐mi , Kang Myung‐Hee , Jang Hae‐Dong , Kwon Young‐In
Published: June 2021

In this study, we investigated the effect of long‐term supplementation of selected medicinal herbs such as Cinnamomum loureirii Nees, Magnolia officinale Rehder et Wils, Brassica oleracea var. capitata L., and Ficus carica L. on activity of intestinal a‐glucosidases and expression of sucrase‐isomaltase complex (SI‐complex) in Sprague‐Dawley (SD) rat model, and also evaluated effect of herbal mixture administration on carbohydrate digestibility by postprandial blood glucose lowering test. Using 4 groups of 11 male SD rats, each group were treated with Cabagin‐S® (positive control) and herbal mixture at low or high concentration given daily for 2 weeks. After 2 weeks administration, postprandial blood glucose lowering effect was evaluated and then small intestine biopsy was collected from each tested rat. To confirm the mechanism of action of herbal administration via regulation of carbohydrate hydrolyzing enzymes, the changes of sucrase/glucoamylase activities and expression of SI‐complex in small intestine were determined. Herbs have regulatory effects on both activity and expression of carbohydrate hydrolyzing enzymes in a similar manner with a known anti‐gastroenteric disorder drug, Cabagin‐S®. Furthermore, our findings confirm previous in vitro observations that herbs have inhibitory effect on carbohydrate hydrolyzing enzymes, namely sucrase, maltase, and SI‐complex.
Antioxidant and Antiproliferation Effects of Some Egyptian Food Wastes Extracts on HepG2 Cells
Authors: Awad Khadega , Abdel Nabi Ali , Awney Hala
Published: June 2021

Nutraceutical compounds derived from food industry wastes as a source of natural phenolics are in great demand as a food supplement due to its antioxidant and anticarcinogenic effects. This study aimed to determine content and profile of phenoilc compounds, antioxidant and anticancer effects of some food industry wastes.Methanolic extracts of artichoke bracts (Cynara cardunculus)(AB), date palm raceme (Phoenix dactylifera) (DPR), Prickly pear peel (Opuntiaficus‐indica) (PPP), Peanut (Arachishypogaea L) skin (PS) and shell (PSH) and empty pea pods (Pisumsativum L) (EPP) were prepared. Phenolic content was assayed by Folin‐Ciocalteau reagent. Phenolic profile was identified by LCMSMS and GC MS. Antioxidant activity was measured by DPPH assay whereas in vitro antioxidant effects were assayed on HepG2 cells by measuring SOD and GPx activities. Anticancer activity was determined by detecting the viability of HepG2 cells by MTT assay. Our data demonstrated that a major phenolics were p – Coumaric acid in PS, alpha‐Tocopherol in DPR, rutin in PPP, luteolin in PSH and myrecetin in EPP whereas a variety of phenolic acids were detected in AB. The highest IC50 for free radical scavenging activity was detected in DPR (21.7 μg/ml). The antiproliferation effect on HepG2 cells was only detected with DPR, PPP and EPP. DPR induced the highest enzyme activity for SOD and GPX. Our study give more interest to investigate the effect ofphenolics of some Egyptian food industry wasteespecially DPR as protective supplement in our diet against disease caused by oxidative stress specially cancer.
Anti‐atherosclerotic effect of sinigrin in ApoE‐deficient mice
Authors: Jang Yeon Jeong , Pyo Suhkneung
Published: June 2021

Atherosclerosis is a chronic inflammatory disease initiated by vascular inflammation and monocyte recruitment in part. Sinigrin belongs to the family of glucosides found in some plants of the Brassicaceae family such as Brussels sprouts, broccoli, and the seeds of black mustard (Brassica nigra). Since sinigrin has been shown to have a variety of biological activities including anti‐inflammatory activities, we examined the effect of dietary sinigrin on atherosclerosis. 6 male ApoE –/– mice and 6 male control (C57BL/6J) mice were fed with 30% high‐cholesterol diet for 21 weeks. 6 of the ApoE –/– mice were also given sinigrin (oral administration of 25 μg /kg body weight, every two days for 21 weeks). Sinigrin had significant suppressive effects on the expression of VCAM‐1 and ICAM‐1 in ApoE –/– mice. In addition, sinigrin attenuated the level of oxLDL, HDL, LDH, triglyceride and total cholesterol in serum. The serum levels of sterol‐regulatory element binding protein‐2 (SREBP‐2), oxidized low‐density lipoprotein receptor‐1 (LOX‐1) and Liver X receptors (LXRs) were inhibited by sinigrin. Furthermore, the treatment of sinigrin reduced the serum levels of IL‐6 and TNF‐α. Taken together, these results suggest that sinigrin has a protective effect against atherosclerosis.
Wnt5a Signaling in Atherosclerosis, its Effect on OxLDL Uptake and Foam Cell Differentiation
Authors: Ackers Ian , Szymanski Candice , Duckett K Jordan , McCall Kelly , Malgor Ramiro
Published: June 2021

Atherosclerosis is a chronic inflammatory disease involving plaque buildup in the arteries and can lead to heart attack and stroke. A key step in the pathogenesis of atherosclerosis is oxLDL uptake by macrophages. Our lab has shown that oxLDL up regulated transcription of Wnt5a in macrophages. Wnt5a and Frizzled 5 (Fz5), a Wnt5a receptor, are highly expressed in foam cells in atherosclerosis. Our hypothesis is that Wnt5a stimulates oxLDL uptake and that macrophages show differential expression of Wnt5a receptors.

In vitro studies: THP‐1 cell line was cultured and differentiated into macrophages. Cells were treated with oxLDL, nLDL, and rWnt5a in combinations to determine their effects on oxLDL uptake. Expression of receptors involved in oxLDL uptake such as CD68, CD36, and scavenger receptor (SR) were analyzed using immunofluorescence, western blot and PCR. To study Wnt5a signaling ROR2, Fz5, and Wnt5a were evaluated in THP‐1 cells as well as in human atherosclerotic tissue samples

In THP‐1 cells oxLDL and Wnt5a increased CD68, CD36, SR and Fz5 at both protein and RNA level. IHC results show that Wnt5a is expressed by macrophages, foam cells, smooth muscle cells (SMC), and endothelial cells. But Fz5 is expressed only by some macrophages, and almost negative in SMC. We also found that ROR2 followed a similar expression pattern to Fz5 in macrophages while SMC near the lipid core were positive for ROR2 but negative for Fz5.

In conclusion, our results suggest that oxLDL, through Wnt5a signaling, plays a role in foam cell formation and oxLDL uptake. In addition, expression of Wnt5a receptors is variable between cell types in atherosclerosis.
Correlates of Renal Atrophy in Murine 2 Kidney 1 Clip Hypertension
Authors: Kashyap Sonu , Ghossan Roba , Zaia Paula , Grande Joseph
Published: June 2021

Renal artery stenosis (RAS) is a common disorder associated with both cardiovascular and renal morbidity and mortality. Recent studies have shown that stenting of the stenotic kidney fails to improve renal or cardiovascular outcomes, prompting a search for more effective therapies. Strain‐specific variability in renal outcomes is a well recognized problem associated with murine models of chronic injury. The objective of this study was to define strain specific differences in renal outcome and correlation with parameters of injury in a murine model of RAS.

We used a 0.25 mm polytetrafluoroethylene cuff to establish RAS in mice (n=292 total). We examined the correlation between age, time following surgery, body weight, blood pressure, heart and kidney weights, ratio of STK/CLK weight, and plasma angiotensin level in 3 murine strains. Percent atrophy in the STK as a function of time following RAS surgery is summarized in the table below:

Strain
1 Week
2 Week
4 Week
6 Week

129
21%
31%
XX
45%

C57BLKS/J
25%
51%
63%
72%

C57BL/6J
XX
XX
39%
41%

The right kidney weight and the STK/CLK weight ratio correlated with %atrophy in all strains, with the strongest correlation observed in C57BL/6J mice. In C57BL/6J mice, blood pressure correlated with %atrophy. CLK weight correlated with heart weight, with the strongest correlation observed in 129Sv mice. In BLKS mice, the time following surgery and age correlated with CLK weight and %atrophy in the STK; blood pressure correlated with plasma angiotensin level.

We conclude that RAS produces severe chronic renal injury in the STK of all strains studied, with the most severe injury observed in C57BLKS/J mice.The STK/CLK weight ratio correlated most strongly with %atrophy and can be used to predict development of chronic renal damage in all strains. Hypertrophy of the CLK, as measured by CLK weight, strongly correlated with heart weight.
Role of PDZ4 Domain of PDZK1 in the Regulation of Cholesterol Metabolism via the Hepatic HDL Receptor SR‐BI
Authors: Kocher Olivier , Birrane Gabriel , Halford Jennifer , Pal Rinku , Sheng Ren , Cho Wonhwa , Penman Marsha , Elowe Nadine , Krieger Monty
Published: June 2021

The four PDZ (PDZ1‐PDZ4) domain‐containing‐adaptor protein PDZK1 controls the expression, localization and function of the HDL receptor SR‐BI in hepatocytes. Many PDZ domains function by binding to the C‐terminus of a target binding partner (canonical binding) and alternatively some PDZ domains directly bind to the inner leaflet of the plasma membrane (membrane anchor). Using transgenic mice expressing internal deletions of selected PDZ domains of PDZK1 in the liver, we found that only PDZ1 and PDZ4 were necessary for PDZK1's full regulation of hepatic SR‐BI. A PDZK1 transgene encoding only the major SR‐BI binding PDZ domain PDZ1 (ΔPDZ2/ΔPDZ3/ΔPDZ4) exhibited dominant negative activity when expressed in WT mice, resulting in the high plasma cholesterol and reduced hepatocyte membrane expression of SR‐BI and PDZK1, seen in PDZK1 KO mice. Unlike full‐length PDZK1, the ΔPDZ2/ΔPDZ3/ΔPDZ4 mutant failed to compensate for the absence of PDZK1 and reduced SR‐BI activity in PDZK1 KO mice. In contrast, a PDZK1 hepatic transgene encoding both the PDZ1 and PDZ4 domains, but not PDZ2 or PDZ3 (ΔPDZ2/ΔPDZ3), was not dominant negative in WT mice, but complemented almost fully hepatic SR‐BI defects in PDZK1 KO mice. Our results show that the PDZ1 and PDZ4 domains together are the only PDZ domains of PDZK1 that are necessary for the normal biological function of hepatic SR‐BI. The PDZ1 domain participates in canonical binding to the C‐terminus of SR‐BI and the PDZ4 domain apparently anchors PDZK1 to the plasma membrane.
Oxidative Damage is Correlated with Mitochondrial Autophagy
Authors: Perry George , Perry Elizabeth , Moreira Paula , Correia Sonia , Castellani Rudy , Wang Xinglong , Lee Hyoung‐gon , Zhu Xiongwei
Published: June 2021

Alzheimer disease (AD) and aging are marked by oxidative damage and mitochondrial abnormalities. Since mitochondria can play a critical role in oxidative damage, we conducted this study to identify the relationship of oxidized RNA, 8‐hydroxyguanosine (8OHG), and mitochondrial DNA (mtDNA) accumulation in AD and aging individuals. Abnormalities were examined by using densitometry of hippocampal pyramidal neurons: mtDNA accumulation as a marker of mitophagy and oxidative damage by 8OHG. Among aging individuals, oxidative damage and mtDNA were highly correlated (r2 = 0.86). While both 8OHG and mtDNA were at higher levels in AD individuals, they were uncorrelated (r2 = 0.06). In contrast, as we found before, oxidative damage was inversely correlated with amyloid‐β; it was unrelated in normal aging individuals. These results suggest that oxidative damage is directly related to mitophagy in aging individuals. With the onset of AD, amyloid‐β plays a strong antioxidant role. These findings indicate that the onset of AD is marked by a pleotrophic change in oxidative stress, one characterized by a change from mitochondria to amyloid‐β dependency.
Cyclin A2 Promotes DNA Repair and Neural Stem Cell Growth
Authors: Gygli Patrick , Chang Joshua , Gokozan Hamza , Catacutan Fay , Schmidt Theresa , Griveau Amelie , Michowski Wojciech , Wong Michael , Palanichamy Kamalakannan , Sicinski Peter , Czeisler Catherine , Otero Jose
Published: June 2021

Mutations in DNA repair genes show peculiar defects in central nervous system morphogenesis characterized by dramatic cerebellar dysmorphia with relatively preserved forebrain development. Loss of the S‐phase cyclin CCNA2 parallels the phenotype of other DNA repair pathway genes. We utilized theoretical biology approaches based on the literature and our own data to derive mechanistic insights to the growth dynamics of the forebrain and cerebellar stem cell niches. We discovered that deletion of CCNA2 delays early embryonic forebrain expansion, which is compensated by increased proliferation at later ages. These data indicate that VZ/SVZ growth is controlled in part due to a proliferative auto‐inhibition, which we utilize in a new mathematical model for VZ/SVZ growth focusing on the period after E14.5. Our data indicate that the growth dynamics of the forebrain niche are not significantly driven by progenitor cell apoptosis relative to the cerebellar niche. Supporting our initial notion that CCNA2 is implicated in DNA repair are our novel findings that Cyclin A2 protein localizes to foci of γH2AX, it is a novel early sensor of DNA double strand breaks lying upstream of H2AX phosphorylation, and CCNA2 loss sensitizes cells to ionizing radiation and may serve as a novel radiation sensitizing target for cancer therapy.
Modeling Human MEGF10 Myopathy in Drosophila melanogaster
Authors: Matin Bahar , Blomquist Hannah , Chen Ci , Kopin Alan , Kang Peter , Draper Isabelle
Published: June 2021

Mutations in MEGF10 underlie a rare congenital muscle disease in humans. MEGF10 is a single transmembrane receptor that is expressed in skeletal muscles as well as in CNS glial cells, in both mammals and Drosophila. Expression of Drpr, the Drosophila homolog of human MEGF10, in fly muscles offers a unique opportunity to probe the role of this receptor in muscle physiology and disease. We have shown that loss‐of‐function mutations in Drpr lead to muscle alterations that recapitulate important features of the human disease. To complement this genetic tool, we have now developed a MEGF10 gain‐of‐function model in Drosophila. In the present study we show that ubiquitous or skeletal muscle‐specific overexpression of either Drosophila Drpr, or mouse MEGF10, in flies leads to decreased viability, whereas heart or glia‐specific overexpression does not. Preliminary analysis carried out by overexpressing the different mammalian or fly MEGF10 receptor orthologs, suggests an important role for the conserved extracellular EMI domain of the protein, to ensure normal function in muscle. Our complementary MEGF10 loss of function/gain of function Drosophila models enable us to begin dissecting the conserved functional pathways regulated by this protein in muscle cells. Together our studies indicate that narrow alterations in the levels of MEGF10 expression may impair normal muscle physiology in vivo. This study was funded by NIHgrant R01 NS080929 (PBK and ID).
Effects of GAT107, an Allosteric Agonist and Positive Modulator of α7 Nicotinic Acetylcholine Receptors, on Chronic Inflammatory and Neuropathic Pain in Mice
Authors: Bagdas Deniz , Wilkerson Jenny , Lichtman Aron , Thakur Ganesh , Damaj M Imad
Published: June 2021

Agonists and positive allosteric modulators (PAMs) of α7 nicotinic acetylcholine receptors (nAChRs) are currently being considered as novel therapeutic approaches for pain modulation. Recent discovery of compounds that can function as dual allosteric agonists and PAMs (ago‐PAMs) added a new pharmacological approach for the regulation of α7 nAChR function.

GAT107 [(+)‐enantiomer of racemic 4‐(4‐bromophenyl)‐3a,4,5,9b‐tetrahydro‐3H‐cyclopenta[c]quinoline‐8‐sulfonamide] is such an agent which has been shown to be an α7‐selective ago‐PAM.

The present study investigated whether GAT107 attenuates chronic inflammatory and neuropathic pain in mice. We found that systemic administration of GAT107 significantly reversed pain behaviors in formalin test, complete Freud adjuvant (CFA)‐ and lipopolysaccharide‐induced inflammatory pain, chronic constriction injury‐induced neuropathic pain, without motor impairment. GAT107 failed to show antinociceptive effects on acute thermal stimulus. The antinociceptive effects of GAT107 were mediated by of α7 nAChRs. Our sites studies suggest that spinal but not peripheral sites mediate in the effect of GAT107 in CFA model. Finally, GAT107 blocked pain‐induced aversion after acetic acid injection, using a conditioned place aversion test. The present results suggest that α7 nAChR modulation by ago‐PAMs may provide new targets in inflammatory and neuropathic pain.

This work was supported by VCU Massey Cancer Center, TUBITAK (2219).
Ocimum basilicum essential oil complexed with β‐Cyclodextrin increases fos protein expression in CNS areas and produces analgesic profile
Authors: Quintans Lucindo , Nascimento Simone , Araujo Adriano , De Lucca Waldecy , Antoniolli Angelo , Quintans Jullyana , Bonjardim Leonardo
Published: June 2021

Ocimum basilicum leaves essential oil (LEO) is rich in monoterpenes, such as linalool and geraniol, and it have shown analgesic effects, but short half‐life and water insolubility are restraint for LEO therapeutic approaches. β‐cyclodextrin (β‐CD) has been applied to improve pharmacological profiles of essential oils or related compounds. Thus, we assessed the antihiperalgesic effect of LEO, alone, or complexed in β‐CD (LEO/β‐CD), on chronic non‐inflammatory muscle pain (an animal model for fibromyalgia) and evaluate the c‐Fos expression in the mice CNS). Mice were treated every day with either LEO/β‐CD (25, 50 or 100 mg/kg, p.o.), LEO (25 mg /kg, p.o.), tramadol (TRM 4mg /kg, i.p.) or vehicle (saline), and 60 minutes after treatment parameters were assessed. Oral treatment with LEO or LEO‐βCD, at all doses tested, produced a significant reduction of mechanical hyperalgesia (p<0.001) and they are able to significantly (p<0.05) increase Fos protein expression in nucleus raphe magnus and periaqueductal gray. Our results provide evidence suggesting that LEO and LEO/β‐CD produces analgesic effects through an activation of CNS areas, such as periaqueductal grey.

Financial Support: CNPq/Brazil, CAPES/Brazil, FAPITEC/SE/Brazil and FINEP/Brazil
B Cell‐Targeted Therapy with Anti‐CD20 Monoclonal Antibody Reduced Secondary Tissue Damage and Enhanced Behavioral Recovery Following Experimental Spinal Cord Injury in Mice
Authors: Cuzzocrea Salvatore , Impellizzeri Daniela , Bruschetta Giuseppe , Cordaro Marika , Crupi Rosalia , Esposito Emanuela
Published: June 2021

Traumatic injury to the spinal cord activates B cells, which culminates in the synthesis of autoantibodies. The functional significance of this immune response is unclear. Antibodies produced after SCI caused pathology, in part by activating intraspinal complement and cells bearing Fc receptors. These data indicate that B cells, through the production of antibodies, affect pathology in SCI. There is increasing appreciation of the important role of B cells in spinal cord trauma and consequently, increasing interest in treating these disorders through B cell‐depletion therapy. The purpose of this study was to investigate the effects of anti‐CD20 mAb B cell depletion therapy within the first 24 hours of SCI.

In this study, the T6‐T7 vertebrae in three groups of mice were injured by a 50‐g clip‐induced compression method, and the anti‐murine CD20 IgG2a antibody (clone 18B12, 1 mg/kg) was intravenously administered starting 1 hour and 6 hours postinjury. Animals were sacrified at 24 hours. The anti‐CD20 antibody treatment caused significant attenuation of leukocyte infiltration, reactive oxygen species‐associated enzymes, and secondary tissue damage. Basso‐Beattie‐Bresnahan (BBB) scores were significantly higher in anti‐CD20‐treated mice than controls 10 days postinjury. Our data demonstrate an important role of B cells which could possibly lead to B cell‐based strategies for the treatment of SCI.
Small Molecule Inhibitors of Constitutively Active Gsa
Authors: Pals‐Rylaarsdam Robin , Carroll Julie , Ceja‐Ruiz Victor , Turcic Kyle , Jenkins Evan , Habbal Renee , Raub Matthew , Cruz Joseph , Walters D Eric
Published: June 2021

Constitutive activation of Gsα is seen in the genetic disorder McCune‐Albright Syndrome, as well as some endocrine adenomas and in tissues affected by cholera toxin. Random mutagenesis of constitutively active Gα subunits in yeast identified three sites that, when mutated, suppressed the constitutive activity caused by R201H: F142, R231, and L266. Because R231 is on the surface of the protein, near the GTP‐binding site it was selected as a candidate site for drug targeting, with the goal of developing inhibitors of the constitutively active Gs alleles. Docking molecules in silico to the R231 binding pocket on the surface of Gs identified commercially available molecules with high potential to bind to the suppressor site. Eighteen molecules with high predicted binding affinity were tested for the ability to decrease the elevation in basal cAMP observed in HEK cells overexpressing the constitutively active Gs‐R201H allele. Two molecules, 6‐oxo‐N‐{[2‐(pyrrolidin‐1‐yl)pyridin‐4‐yl]methyl}‐1,6‐dihydropyridazine‐3‐carboxamide and N‐{[3‐(3‐methylphenyl)phenyl]methyl}‐6‐oxo‐1,6‐dihydropyridazine‐3‐carboxamide were able to significantly reduce cAMP levels by at least 10% from baseline. Neither of these molecules significantly reduced basal cAMP levels in cells transfected with wild‐type Gs constructs. These molecules will form the basis for further studies to develop antagonists of mutant Gs proteins associated with human disease.

This work was Supported by NIH grant 1R15ED020190‐01
Developmental Changes in DNA Methylation of CYP3A5 in Human Liver
Authors: Vyhlidal Carrie , Bi Charlie , Gaedigk Roger , Ye Shui , Kingsmore Stephen , Leeder J.
Published: June 2021

CYP3A5 is polymorphically expressed in human prenatal and postnatal liver due to the presence or absence of the CYP3A5*3 allelic variant. However, expression of members of the CYP3A subfamily recently has been reported to be regulated through DNA methylation. The object of this study was to examine changes in CpG methylation of the CYP3A5 gene in human pediatric and prenatal liver. Methylation status of CpG dinucleotides was initially determined in 16 pediatric livers using Agilent SureSelect Human Methyl‐Seq and data stratified by age into four groups (20‐96 days; 2‐2.6 yrs; 7‐8 yrs; and 15‐17 yrs). Bisulfite sequencing was performed on 48 pediatric and 34 prenatal livers to confirm results of Methyl‐Seq analysis. Cytosines in the promoter and introns 1 and 7 of CYP3A5 exhibited statistically significant changes in methylation with respect to age group in pediatric livers (p‐values=0.02, 0.03 and 0.02, respectively). Bisulfite sequencing of the CYP3A5 promoter revealed hypermethylation of cytosines at ‐411 and ‐460 in prenatal versus pediatric liver (60% vs 38% and 80% vs 57%, respectively, p<0.0001) which exhibited age‐dependent demethylation during the first year of life (r2=0.85, p<0.0001 and r2=0.53, p=0.005, respectively). Methylation of cytosines at ‐411 and ‐460 was associated with total CYP3A5 mRNA expression (p<0.0001). Current studies are investigating interactions between genotype and methylation to influence CYP3A5 gene expression.
Fibroblast Growth Factor (Fgf) 21 is a Novel Target Gene of Glucocorticoid Receptor
Authors: Vispute Saurabh , Bu Pengli , Le Yuan , Cheng Xingguo
Published: June 2021

Fibroblast growth factor (Fgf) 21 is a metabolic regulator that plays an important role in the maintenance of glucose, lipid, and energy homeostasis. Recent studies also suggest that Fgf21 induction can attenuate chemical‐induced toxicities, such as of acetaminophen, alcohol and dioxin. Glucocorticoid receptor (GR) is indispensable for cell growth and development, drug metabolism as well as immune response. However, the impact of GR activation on regulation of Fgf21 is unknown. The present study was designed to investigate whether activation of GR by the synthetic glucocorticoid Dexamethasone (DEX), can alter Fgf21 expression. Our results showed that in mouse liver, both low (2mg/kg) and high (50mg/kg) doses of DEX up‐regulated Fgf21 mRNA and protein expression. In addition, DEX induced Fgf21 mRNA and protein expression in both mouse (Hepa1c1c7) and human (Hep3B) hepatoma cells in a concentration‐ and time‐dependent manner. Generally, DEX produces its effects via activation of GR and/or pregnane X receptor (PXR). By using PXR‐null mouse model, we showed that 50mg/kg of DEX induced Fgf21 mRNA expression similarly in both wild‐type and PXR‐null mouse liver, indicating that the induction of Fgf21 by DEX is PXR‐independent. The in silico DNA sequence analysis revealed the presence of several putative GR response elements (GREs) in the 3kb promoter of mouse and human Fgf21/FGF21 genes. Further, GR silencing using GR‐specific siRNA, attenuated DEX‐induced Fgf21 expression in both mouse (Hepa1c1c7) and human (Hep3B) hepatoma cells. In summary, activation of GR induced Fgf21 expression in mouse liver as well as in cultured mouse and human hepatocytes.
Therapeutic efficacy of cerium oxide nanoparticles against sepsis induced acute lung injury
Authors: Arvapalli Ravikumar , Manne Nandini , Rice Kevin , Blough Eric
Published: June 2021

Sepsis induced acute lung injury (ALI) is one of the leading causes of mortality in the intensive care unit. The development of ALI is primarily driven by the systemic inflammatory response syndrome and the increased cellular ROS subsequent to immune system over activation. Cerium oxide nanoparticles (CeO2) nanoparticles have been shown to scavenge reactive oxygen species both in vitro and in vivo under various experimental settings while others have suggested that they may also exhibit the ability to protect lungs from hypoxia induced oxidative stress. In the present study we investigated the protective effects of cerium oxide nanoparticles against sepsis induced ALI. Administration of a single dose of CeO2 nanoparticles (0.5mg/kg) to septic animals attenuated alveolar congestion. Treatment associated decreases in alveolar congestion were associated with reduced infiltration of inflammatory cells, diminished myeloperoxidase activity, and decreased p38‐MAPK / STAT3 phosphorylation. Additional experiments to explore the potential efficacy of using CeO2 nanoparticles for the treatment of sepsis induced ALI may be warranted.
The Evaluation of Bac7 Labeled Nanocarriers to Deliver Antiretrovirals as a Pre‐Exposure Prophylaxis (PrEP) Therapy for HIV Prevention
Authors: Nelson Antoinette , Adler Derek , Myers Dan , Szekely Zoltan , Zhang Xiaoping , Sinko Patrick
Published: June 2021

Objective

The objective of this study is to engineer nanocarriers (NCs) capable of translocating through colorectal mucus and tissue and delivering drugs into cells of the distal colon. NCs will be delivered in quickbreak foams to provide sufficient colorectal coverage and be used as a colorectal PrEP therapy to prevent HIV transmission.

Methods

FITC‐labeled poly(ethylene glycol) NCs were conjugated to the protease inhibitor amprenavir (APF) or cell‐penetrating peptide (CPP) Bac7 (BPF). APF and BPF were used to treat Caco‐2 cells in vitro and cell uptake was studied via flow cytometry. Mice were also treated with enemas containing APF or BPF and tissue retention was investigated. Foam formulations were studied for their ability to distribute NCs evenly throughout the distal colon. Histology and toxicity studies were also completed to assess the safety of the foam formulation.

Results

BPF uptake was shown to be 4‐fold higher than APF in vitro. Similar results were seen in vivo and BPF was retained within colorectal tissue for up to 5 days suggesting transport into the lamina propria. Ex vivo tissue samples show complete coverage of the distal colon by foams and NCs. Preliminary toxicity data of foam components display promising results based on histological observations.

Conclusion

Bac7 CPP increased the translocation of NCs across the colorectal mucus and increased uptake into colonic cells both in vitro and in vivo. The current foam formulation is effective in providing uniform coverage of the distal colon and causes minimal tissue damage.

Acknowledgements

Funding by NIH Grants T32GM8339, 1R01AI084137 and R37AI/DK‐051214
Ethanol Discontinuation Does Not Fully Reverse Its Chronic Effects on Blood Pressure, Heart Rate, and Their Variability in SHRs: Diurnally‐Unrelated Effects
Authors: El‐Mas Mahmoud , Abdel‐Rahman Abdel
Published: June 2021

Chronic ethanol alters diurnal hemodynamic and autonomic rhythms in SHRs, but not in, Wistar‐Kyoto rats (WKY). In this study, we investigated if these effects of ethanol could be reversed following ethanol withdrawal. Telemetered SHRs and WKY rats were fed a liquid diet containing 5% w/v ethanol or isocaloric diet for 12 weeks, after which ethanol diet was replaced by control diet for another 2 weeks. Blood pressure (BP), heart rate (HR), hemodynamic variability, and their circadian rhythms were monitored during the last week of ethanol regimen and after withdrawal. Compared to little effects in WKYs, ethanol‐treated SHRs exhibited significant increases in HR and decreases in BP and time‐domain variability of mean arterial pressure (SDMAP) and HR (standard deviation of R‐R intervals, SDRR, and root mean square of successive R‐R differences, rMSSD). Ethanol discontinuation fully and partially reversed the cardiac and hypotensive responses, respectively, but had no effect on the SDMAP reductions caused by ethanol. Unlike WKYs, SHRs showed no diurnal variations in BP or rMSSD, and reversed HR and SDMAP rhythms. The defective diurnal profiles in SHRs continued during ethanol treatment. Further, the hemodynamic profiles following ethanol discontinuation were similar during light and dark cycles. Overall, ethanol discontinuation reverses the cardiac, but not BP, responses in a diurnally independent manner.