bluuebunny/crossref_metadata_2025 huggingface dataset:
Usual Intakes of Flavonoids: Estimates from What We Eat in America, NHANES 2007–2010
Authors: Sebastian Rhonda S , Goldman Joseph D , Enns Cecilia Wilkinson , Moshfegh Alanna J
Published: November 2023

Usual intakes are estimates of the long‐term average daily consumption of foods, food components, or nutrients. Despite a vast number of studies suggesting favorable effects of dietary flavonoids on human health, usual intakes of flavonoids have never before (to our knowledge) been determined. Recent release of a comprehensive database of the flavonoid content of foods has made possible the calculation of usual flavonoid intakes. The purpose of this study was to ascertain and describe distributions of usual intakes of flavonoids (total and 6 classes) in the U.S. population. Intake data collected in What We Eat in America, NHANES 2007–2010 from 17,892 individuals were analyzed. Day 1 and day 2 flavonoid intakes were estimated using Flavonoid Values for USDA Survey Foods and Beverages 2007–2010. For anthocyanidins, flavan‐3‐ols, flavanones, flavones, and isoflavones, the two‐part National Cancer Institute approach for estimating the probability of consumption on a day and the amount consumed (which assumes a correlation between the two) was used. For total flavonoids and flavonols, it was assumed that at least a minimal amount is consumed every day by everyone (i.e., probability of consumption = 1). For individuals age ≥1 year, median usual intake of total flavonoids was 117.18 mg/day. Estimates at the 5th, 25th, 75th, and 95th percentiles were 17.01 mg, 55.37 mg, 236.09 mg, and 595.26 mg, respectively, indicating a positively skewed distribution. Due to high concentrations of flavonoids in certain foods, positively skewed distributions were also observed for all flavonoid classes. In descending order by weight, usual median intake of flavan‐3‐ols was 57.76 mg; flavonols, 14.38 mg; flavanones, 7.02 mg; anthocyanidins, 6.70 mg; flavones, 0.62 mg; and isoflavones, 0.17 mg. On an average day in 2007–2010, the likelihood of consuming the episodically consumed flavonoid classes (i.e., anthocyanidins, flavan‐3‐ols, flavanones, flavones, and isoflavones) ranged from 94% for flavan‐3‐ols to 39% for isoflavones. Tables describing usual intakes of flavonoids (total, class, and 29 individual flavonoids) by selected sociodemographic variables will become available in 2017 on the Food Surveys Research Group web site (www.usda.gov/nea/bhnrc/fsrg). This novel information will add to the body of knowledge concerning flavonoid intake and thus facilitate the eventual establishment of recommendations regarding safe and adequate dietary intakes of these beneficial compounds.

Support or Funding Information

ARS, USDA
The Association between Dietary Fiber Intake and CRP levels, US Adults, 2007–2010
Authors: Nielsen Samara Joy Joy , Trak‐Fellermeier Maria Angélica , Joshipura Kaumudi
Published: November 2023

Background

The 2015 Dietary Guidelines for Americans Committee highlight dietary fiber as one of the four nutrients that Americans under‐consumed. Data from the National Health and Nutrition Examination Survey 1999–2000 on dietary fiber demonstrated a strong inverse relation between the level of dietary fiber intake and C‐reactive protein, but did not analyze specific sources nor the strength of the effect according to the dietary fiber source.

Objective

To determine the relationship between C‐reactive protein (CRP) levels and dietary fiber sources in the US adult population (≥ 20 years of age).

Design

In the 2007–2010 National Health and Nutrition Examination Survey, a cross‐sectional nationally representative sample of the US population, two 24 hour dietary recalls were collected from 9,720 adults 20 years and older. Of those that had both 24 hour recalls, 9,284 adults had CRP data. We categorized CRP into 3 levels, < 0.10 mg/dL, 0.10 mg/dL≤CRP< 0.30 mg/dL, ≥ 0.30 mg/dL. The relationship between CRP levels and total fiber intake as well as the sources of dietary fiber including whole grain intake and fruit and vegetable intake were examined by using multinomial logistic models adjusted for age, sex, race, BMI and smoking.

Results

Adults with lowest quartile of dietary fiber intake compared to the highest quartile were 50% more likely to have higher CRP levels (OR: 1.47; 95% CI: 1.28, 1.68). Similarly, the lowest quartile of whole grain intake compared to the highest quartile was 40% more likely to have higher CRP levels (OR: 1.40; 95% CI: 1.19, 1.65). In addition, the lowest quartile of fiber rich fruit and vegetable intake compared to the highest quartile was 50% more likely to have higher CRP levels (OR: 1.51; 95% CI: 1.27, 1.79).

Conclusions

We found an inverse relationship between CRP levels and dietary fiber consumption. High whole grain, and high fruit and vegetable intake were associated with low C‐reactive protein levels, a marker of systemic inflammation.
Monetary Diet Cost is Positively Associated with Diet Quality: Analysis of School‐Aged Children in South China
Authors: Zhang Jieyi , Xue Hongmei , Li Danting , Yin Jun , Chen Yuanyuan , Cheng Guo
Published: November 2023

Introduction

The association of diet quality with diet cost has been investigated in many developed countries. However, pertinent studies remain scarce in China. As food prices and dietary habits vary with different countries, we aimed to examine the relation of diet quality to monetary diet cost among Chinese Children.

Methods

Data on 1,605 Chinese Children (48.4% girls) aged 7–15 years were analyzed. Dietary data from 3‐d 24h dietary recalls were used to calculate the published Chinese Children Dietary Index (CCDI), which developed based on Chinese Dietary Intake recommendations. CCDI incorporated 16 components (11 foods/food groups, 3 nutrients and 2 health‐promoting behaviors), with a higher score indicating a better diet quality. Monetary costs of diet were calculated by multiplying the average amount of consumed food (g/day) by price of each food and summing the products. The average annual price of each food item was obtained from the government website, supermarkets and farmers' markets, or fast food chains. All food prices, adjusted for preparation and waste using China Food Composition database, were expressed in RMB (¥) per 100g edible portion. As the monetary cost of diet was observed strongly associated with energy intake (r=0.69), monetary diet cost per 1,000 kcal of energy intake per day was calculated. Multivariate linear generalized regression models were used to explore the relevance of diet quality to monetary diet cost.

Results

Subjects spent more money per 1,000 kcal had a higher diet quality indicated by higher CCDI score after adjustment for age, gender, family location, parental educational level, family income level and interaction of age and CCDI score (¥12.35 per 1,000 kcal for the highest‐CCDI‐score quintile vs ¥11.16 per 1,000 kcal for the lowest‐CCDI‐score quintile, p=0.0006). With the improvement of 1 point in CCDI score, the monetary diet costs increased ¥0.14 per 1,000 kcal.

Conclusion

Diet quality is positively associated with energy‐adjusted diet cost among Chinese school‐aged Children, warranting that the public policy for healthful food promotion should totally take economic issues into account.

Support or Funding Information

All phases of this study were supported by research grant from the National Nature Science Foundation of China (No. 81472976)
Dyslipidemia among School‐age Children and Adolescents in Southern Provinces of China
Authors: Shi Jiachen , Liu Liping , Li Hong , Du Shufa
Published: November 2023

Background

Many studies have reported dyslipidemia among adults, while few studies explored dyslipidemia among school‐age children.

Methods

We drew fasting blood samples from 1,834 children and adolescents (1,030 boys and 804 girls) aged 6 to 15 years old in southern provinces of China and measured lipid profiles with Hitachi 7600 biochemistry auto‐analyzer.

Results

The results showed that total cholesterol (TC) was 3.60±0.59 mmol/L (means ± standard deviation, SD); triglyceride (TG) 0.90±0.43 mmol/L; high density lipoprotein (HDL) 1.51±0.36 mmol/L; and low density lipoprotein (LDL) 1.92±0.57 mmol/L. The serum concentrations of TC, TG, HDL, and LDL were higher among girls than boys, but the differences were not statistically significant (p > 0.05). The prevalence of elevated TC, TG and LDL were 2.8%, 8.5%, and 4.5% among those school‐age children, respectively. On average, 10.9% of boys and 11.1% of girls had dyslipidemia.

Conclusion

Dyslipidemia should not be overlooked among school‐age children and adolescents in China.

Support or Funding Information

This study is supported by the Beijing CDC.
The Impact of Gestational Endocrine Disorders of Breast Milk Fat Characteristics and Infant Growth
Authors: Ellsworth Lindsay , Harman Emma , Anderson Clare , Craig Annmarie , John Elizabeth , Yeh Joanna , Gregg Brigid
Published: November 2023

Background

Maternal endocrine disorders are increasingly prevalent co‐morbidities during pregnancy with the potential to impact maternal health, fetal development, newborn health and long‐term childhood outcomes. Limited data exists on changes in breast milk composition associated with maternal metabolic disease in pregnancy. Based on existing data from animal models, we hypothesize that maternal alterations in insulin signaling leads to changes in milk fat content and fatty acid profile with an increased ratio of omega‐6 long chain polyunsaturated fatty acids to omega‐3 long chain polyunsaturated fatty acids and reduced monounsaturated fatty acid content.

Objective

Our study objective is to determine if maternal metabolic disease leads to changes in milk composition that then impacts infant growth trajectory.

Methods

We are in the enrollment phase of a large prospective cohort study of mothers with gestational diabetes mellitus (GDM), type 1 diabetes mellitus, type 2 diabetes mellitus, obesity, and polycystic ovary syndrome (PCOS) in comparison to healthy mothers. This study has been approved by the University of Michigan Institutional Review Board. We will perform metabolomic analysis on human milk to evaluate fatty acid content through mass spectrometry. We will evaluate mammary epithelial cells for protein and mRNA levels of lipogenic enzymes to correlate to milk fat composition. We will determine if maternal factors including metabolic disease diagnosis, body mass index, and glycemic control influence milk composition. We will then correlate infant growth trajectory with milk fat alterations.

Results

We have enrolled 29 mother‐baby pairs including 16 healthy, 6 obese, 5 PCOS, and 2 GDM mothers. Mothers had a mean maternal age of 30 years old, mean gestation age of 40 weeks, and 31% delivered by C‐section. In our existing cohort, growth from birth to 2 months does not reveal a significant difference in weight or length however there is an indication that infant head circumference may differ by maternal metabolic disease category by 2‐way ANOVA testing. Breast milk composition analysis is currently underway.

Conclusions

Additional recruitment is ongoing with the goal of expansion of the diabetes mellitus cohorts in order to evaluate for significant differences in breast milk fat composition and infant growth trajectory in the setting of maternal metabolic disease.

Support or Funding Information

Percy J. Murphy, M.D. and Mary C. Murphy, R.N. Endowed Children's Research Fund
Concentration of ghrelin and leptin in human serum and human milk in exclusively breastfeeding nursing mothers. Preliminary data
Authors: Vasquez‐Garibay Edgar M , Larrosa‐Haro Alfredo , García‐Arellano Samuel , Guzmán‐Mercado Elizabeth , Muñoz‐Esparza Nelly , Muñoz‐Valle Francisco , Romero‐Velarde Enrique
Published: November 2023

Introduction

Leptin and ghrelin are two key hormones in the regulation of hunger and appetite that are present in human milk and an association between serum and human milk has been demonstrated between these two hormones with anthropometric indicators and body composition in three‐ and six‐month‐old infants. Therefore, the purpose of this report is to show the relationship and difference between serum and human milk concentration of ghrelin and leptin in nursing mothers who exclusively breastfeed (EBF) to four‐month‐old infants.

Methods

Serum and human milk concentrations of ghrelin (n = 11) and leptin (n = 16) were determined in nursing mothers of EBF four‐month‐old infants attended at the Nuevo Hospital Civil de Guadalajara. Fasting blood samples were obtained in Vacutainer® tubes with anticoagulant EDTA treated with protease inhibitors aprotinin and dipeptidyl peptidase‐4 inhibitor. Human milk was collected from fasting nursing mothers before and after breastfed their infants. ELISA methods were used to determine the levels of leptin and ghrelin in the serum and milk samples. (Leptin Ultrasensitive, ALPCO® USA and Human Ghrelin Total, EMD Millipore, Billerica, USA, respectively). The comparison and the relationship between serum and human milk concentration of the hormonal biomarkers were performed by Mann‐Whitney U tests and Pearson's correlation coefficient. The protocol was approved by the Research and Bioethics Committee of the New Civil Hospital of Guadalajara.

Results

Serum levels of ghrelin (501 pg/mL) were significantly greater than in human milk (138 pg/mL) and serum levels of leptin (9.8 ng/mL) were also greater than in human milk (0.4 ng/mL), p<0.001. The concentration of ghrelin and leptin in foremilk and hind milk showed no significant differences. There was a high correlation between leptin concentration in serum and human milk (r=0.881, p<0001).

Conclusions

Concentrations of these appetite regulating hormones in the mother's serum were greater than in their own milk. The high correlation between serum and milk leptin concentration in nursing mothers of EBF infants suggests that the variation in leptin concentration in human milk is probably explained by the serum concentration of maternal leptin.

Support or Funding Information

Project supported by the National Council of Science and Technology of Mexico (234158)
Indoxyl Sulfate Involves Abnormality of Iron Metabolism Through Hepcidin Regulation
Authors: Hamano Hirofumi , Ikeda Yasumasa , Watanabe Hiroaki , Horinouchi Yuya , Izawa‐Ishizawa Yuki , Ishizawa Keisuke , Tsuchiya Koichiro , Tamaki Toshiaki
Published: November 2023

Background

Hepcidin, a secreted hormone derived from hepatocytes, a key regulator of iron metabolism, is shown to be elevated in chronic kidney disease (CKD), leading to the dysregulation of iron metabolism in CKD. Indoxyl sulfate (IS), a uremic toxin, is accumulated during the progression of CKD, and it exacerbates CKD through oxidative stress and inflammation. However, the role of IS on iron metabolism has remained unclear. In the present study, we investigated the involvement of IS on iron metabolism.

Methods

In in vitro, HepG2 cells were used to examine IS effect on hepcidin. In in vivo, we analyzed hepcidin, iron transporters, and tissue iron content by using a mice model of CKD. In addition, we investigated whether CKD‐induced iron dysmetabolism was ameliorated by AST‐120, uremic toxin adsorbent.

Results

IS stimulation augmented hepcidin expression in a dose‐dependent manner in HepG2 cells. IS‐mediated hepcidin upregulation was inhibited by silencing of aryl hydrocarbon receptor (AhR), a receptor of IS. IS also augmented oxidative stress and hepcidin upregulation with IS was suppressed by an anti‐oxidant agent. CKD mice with increased IS accumulation showed the elevation of hepatic hepcidin expression and blood hepcidin concentration compared with control mice. In CKD mice, ferroportin expression was changed in duodenum and spleen, and splenic iron content was increased. These changes were ameliorated by AST‐120 treatment.

Conclusion

Our study suggested that IS participates in the dysregulation of iron metabolism through hepcidin regulation. Removal of IS from plasma might be a therapeutic strategy for abnormality of iron metabolism in CKD.
Characterization of Nefazodone Time‐dependent Inhibition of Cytochrome P450 3A
Authors: Wang Zhican , Rock Brooke M , Pearson Josh T , Tran Thuy , Rock Dan A
Published: November 2023

Nefazodone (Serzone®), marketed as an antidepressant, was discontinued in 2004 due to the rare incidence of hepatoxicity. In vitro studies focused on characterizing the metabolism and potential bioactivation pathways of nefazodone revealed that metabolism of nefazodone by CYP3A leads to the formation of multiple reactive metabolites that could contribute to nefazodone toxicity (Argoti et al., 2005; Kalgutkar et al., 2005; Bauman et al., 2008). The literature also reported nefazodone as a time‐dependent inhibitor (TDI) of CYP3A4 (Kalgutkar et al., 2005; Bauman et al., 2008). However, the plausible mechanism of nefazodone‐mediated inactivation of CYP3A4, particularly the role of reactive metabolites in CYP3A4 inactivation, remains unclear. In the work presented here, a comprehensive investigation of the CYP3A4‐ and CYP3A5‐mediated metabolism of nefazodone was conducted to characterize the difference in metabolite formation, enzyme inhibition kinetics and conjugation of reactive metabolites towards metabolism‐mediated inactivation of CYP3A. Nefazodone caused reversible inhibition for both CYP3A4 and CYP3A5, while TDI was observed for only CYP3A4. Studies in human liver microsomes, CYP3A4 and CYP3A5 supersomes indicated the formation of multiple reactive metabolites and sequential conjugation with nucleophilic glutathione (GSH) and/or cyanide. Screening of TDI on CYP3A4 was also applied for two metabolites of nefazodone. Interestingly, the major oxidative nefazodone metabolite (hydroxyl‐nefazodone) also caused a significant TDI effect towards CYP3A4; whereas, 1‐(3‐chlorophenyl) piperazine (m‐CPP) had negligible TDI effect. In addition, the mechanism of TDI was further explored by monitoring carbon monoxide binding, formation of metabolic intermediate complex, and intact heme loss after pre‐incubation of CYP3A4 with nefazodone. Finally, the potential for apo‐adduct formation with CYP3A4 was evaluated. The results support the hypothesis that CYP3A4 inactivation by nefazodone occurs via formation of reactive metabolites that covalently modify CYP3A4.
Treatment with the Soluble Guanylate Cyclase Stimulator IW‐1973 Reduces N‐terminal Pro B‐type Natriuretic Peptide and Improves Heart Function in Rat Models of Heart Failure with Preserved Ejection Fraction and Heart Failure with Reduced Ejection Fraction
Authors: Shea Courtney , Liu Guang , Sarno Renee , Lonie Elisabeth , Zimmer Dan , Currie Mark , Masferrer Jaime
Published: November 2023

Introduction

Intracellular cGMP levels are reduced in many cardiovascular diseases, including heart failure, due to high levels of reactive oxygen species, which inactivate NO, the endogenous ligand of soluble guanylate cyclase (sGC). sGC “stimulators” bind sGC receptors and synergize with NO to increase cGMP production. IW‐1973 is a novel sGC stimulator in clinical development that has been previously shown to lower mean arterial pressure and heart hypertrophy in Dahl Salt‐Sensitive (Dss) rats.

Hypothesis

We assessed the hypothesis that sGC stimulator IW‐1973 would preserve cardiac function in 2 rat models of heart failure: Dss (HFpEF) and post MI (HFrEF).

Methods

In male Dss rats, heart failure was induced by high salt diet (8%); IW‐1973 (10 mg/kg/day in chow; n=8) was added after 2 weeks, for 5 more weeks. In a post MI model, heart failure was induced by surgical ligation of the left coronary artery of male Sprague Dawley rats (n=10), IW‐1973 (10 mg/kg/day in chow) was administered for 8 weeks. In both models, echocardiography was conducted at the end of the experiment; at necropsy, plasma, serum, urine and tissues samples were collected. All comparisons are vs untreated controls (n=9 and 11).

Results

In Dss, IW‐1973‐treated rats had lower heart/body mass ratio (3.8±0.1 vs 4.6±0.1 g/kg, p<0.05), liver/body mass (43.2±1.1 vs 49.2±1.8 g/kg, p<0.05) and albumin creatinine ratio (1.2±0.1 vs 2.4±0.3 g/g creat/kg, p<0.05) vs controls. By echocardiography IW‐1973‐treated rats had higher ejection fraction vs controls (85±2 vs 70±3%, p<0.05). In post MI, IW‐1973‐treated rats had lower left ventricular end systolic area vs controls (32±2 vs 39±2 mm2, p< 0.05). In Dss and post MI, IW‐1973‐treated rats had lower levels of N‐terminal pro B‐type natriuretic peptide (NT‐proBNP) vs controls (271±13 vs 585±98, p<0.05, 274±28 vs 581±81, pg/ml p<0.05, respectively.)

Conclusions

IW‐1973 preserved heart function in rat models of HFpEF and HFrEF, including reducing levels of the clinically relevant heart failure biomarker, NT‐proBNP.
Essential role for interleukin‐12 in microvascular endothelial dysfunction in type 2 diabetes
Authors: Radwan Eman , Mali Vishal , Haddox Samuel , Trebak Mohamed , Souad Belmadani , Matrougui Khalid
Published: November 2023

Introduction

It is well established that type 2 diabetes (T2D) is associated with microvascular dysfunction. However, the in vivo role of interleukin‐12 (IL‐12) in microvascular dysfunction in type 2 diabetes is an important question that remains unanswered.

Methods and Results

In the present study, we utilized C57/Bl6, db−/db−, IL‐12−/−, db−/db−IL‐12−/−, endoplasmic reticulum (ER) stress CHOP−/−, and db−/db−CHOP−/− mice. We measured IL‐12 plasma level using kit assay and determined the microvascular function. We found an increase in IL‐12 plasma level in db−/db− compared to C57/Bl6 indicating the potential injurant effect of IL‐12 in T2D. Microvascular endothelial‐dependent relaxation was impaired in db−/db− compared to C57/Bl6, IL‐12−/−, and ER stress CHOP−/− mice. Interestingly, endothelial‐dependent relaxation was improved in db−/db−IL‐12−/− and db−/db−CHOP−/− mice.

Conclusions

Our data demonstrate that the disruption of IL‐12 rescued the microvascular endothelial function through ER stress CHOP‐dependent mechanism.
Interaction of obesity and Nrf2 deficiency accelerates cerebromicrovascular aging and impairs neurovascular coupling responses
Authors: Valcarcel‐Ares Marta Noa , Tarantini Stefano , Hertelendy Peter , Tucsek Zsuzsanna , Gautam Tripti , Kiss Tamas , De Cabo Rafael , Farkas Eszter , Kinter Micheal , Sonntag William E. , Csiszar Anna , Ungvari Zoltan
Published: November 2023

Moment‐to‐moment adjustment of cerebral blood flow (CBF) via neurovascular coupling has a critical role in maintenance of healthy cognitive function. Previous studies demonstrate that obesity promotes cognitive impairment due to, at least in part, its deleterious effects on neurovascular coupling. Importantly, the effects of obesity are exacerbated in aging, but the underlying mechanisms remain elusive. There is increasing evidence that Nrf2/ARE signaling plays a critical role in vascular resilience to obesity and the metabolic syndrome and that aging is associated with progressive Nrf2 dysfunction, promoting microvascular oxidative stress. The present study was designed to test the hypothesis that Nrf2 deficiency contributes to neurovascular uncoupling and exacerbate cerebromicrovascular dysfunction induced by obesity. To test this hypothesis Nrf2+/+ and Nrf2−/− mice were fed a standard diet or an adipogenic high fat diet (HFD) and cortical neurovascular coupling responses were assessed. In control mice feeding HFD for 6 months significantly increased vascular oxidative stress and decreased endothelium‐mediated CBF responses (laser Doppler flowmetry) in the somatosensory cortex evoked by contralateral whisker stimulation. Nrf2 deficiency significantly exacerbated HFD‐induced neurovascular uncoupling, mimicking the aging phenotype. Together, these results provide additional evidence that intact Nrf2 signaling importantly contributes to cerebromicrovascular health, preserving neurovascular coupling mechanisms. Further studies are needed to elucidate the endothelial and astrocytic mechanisms impacted by Nrf2 deficiency and HFD‐induced obesity, determining the potential roles of oxidative stress and chronic low‐grade neuroinflammation.

Support or Funding Information

This work was supported by grants from the American Heart Association, the National Center for Complementary and Alternative Medicine, and the National Institute on Aging
Angiotensin II‐ mediated vasoconstriction of the visceral adipose tissue vasculature is linked to systemic hypertension in obesity
Authors: Park Song‐Young , Karki Shakun , Saggese Samantha M. , Zuriaga Maria A. , Carmine Brian , Hess Donald , Apovian Caroline , Gokce Noyan
Published: November 2023

Objectives

Central adiposity, in particular the accumulation of intra‐abdominal visceral fat, has been linked to adipose tissue dysfunction and obesity‐related diseases including hypertension and atherosclerosis, however regulatory mechanisms are poorly understood. We hypothesized that increased micro‐vascular tone specifically in visceral fat may contribute to abnormal peripheral vascular resistance.

Methods

In 30 obese subjects(age 42±10 yr, BMI 42±3 kg/m2) undergoing elective bariatric surgery, we characterized vaso‐contractile responses of isolated subcutaneous (SC) and visceral (VIS) adipose tissue arterioles using videomicroscopy, and examined relevant depot‐specific gene expression using qPCR.

Results

Visceral arterioles exhibited significantly greater angiotensin II‐mediated vasoconstriction by 20% in obese hypertensives (OH, SBP 146±8 mmHg) compared to obese normotensives (O, SBP 118±5 mmHg) (p<0.05). Additionally, VIS arterioles showed greater vasoconstriction in response to angiotensin II compared to SC in OH (by 25%, p<0.05) but not in O. Angiotensin converting enzyme (ACE) mRNA expression was significantly increased in VIS compared to SC (2‐fold, p<0.05) in OH but not O. In contrast, vasopressor responses to other agonists including endothelin‐1 and phenylephrine were not different between fat depots or subjects with or without hypertension.

Conclusion

The findings demonstrate differential activation of the renin‐angiotensin system (RAS) in the visceral adipose tissue microvasculature that may contribute to mechanisms of systemic hypertension in obesity. RAS targeting, in particular, may provide incremental clinical benefit in treating obesity‐related hypertension.

Support or Funding Information

NIH grants P01HL081587 and RO1 HL1145675
Neural Stem Cells Reduce Symptomatic Inflammation and Mortality in Aged Stroke Mice following Delayed tPA Treatment
Authors: Eckert Auston D. , Hamblin Milton , Lee Jean‐Pyo
Published: November 2023

Stem cell therapy offers great promise in treating stroke. Extensive injury from stroke results from ischemia‐reperfusion (IR), which damages the blood‐brain barrier (BBB), promotes inflammation and loss of cerebral parenchyma. Presently, tissue plasminogen activator (tPA) is the sole FDA‐approved antithrombotic treatment available for stroke. tPA has a narrow time window (<4.5hour) and delayed tPA (>5h) can exacerbate BBB damage and increase hemorrhagic complications. Our long‐term goal is to limit early‐stage BBB injuries, an outcome that would protect against the second phase of stroke damage. Aging is a strong risk factor for stroke but the harmful effects of delayed tPA (>4.5h), particularly on aged stroke animals, have not been well studied. We utilized a mouse model of stroke with middle cerebral artery occlusion (MCAO) to induce focal ischemia followed by reperfusion. We injected human neural stem cells (hNSCs) 24h after MCAO to correspond to the upregulation of endogenous proinflammatory cytokines. Previously, we reported that intracranial transplantation of hNSCs rapidly ameliorates BBB damage caused by ischemic stroke in young adult mice. In this study, we employed aged mice to test the effect of delayed tPA‐induced IR injury. We tested the hypothesis that hNSCs ameliorate pathophysiology of aged stroke mice after delayed tPA administration (6h post‐MCAO). We found that delayed tPA administration exacerbates BBB damage, upregulates inflammatory cytokines, and increases mortality. Engrafted hNSCs reduced BBB disruption, rapidly ameliorated symptomatic inflammation, and decreased mortality within 24h post‐transplantation (48h post‐MCAO). Our data demonstrate clear benefits of hNSCs during the early stage of ischemic stroke in aged stroke mice.

Support or Funding Information

1. Louisiana Clinical and Translational Science Center (LA CaTs), Pilot Grant, to J‐P. L. 2. Carol Lavin Bernick Faculty Grant to J‐P. L, Tulane University.
Interferon type II increases lipolysis and fatty acid oxidation in pancreatic β cells
Authors: Truong Nguyen , Bazil Jason , Olson L. Karl
Published: November 2023

Type 1 diabetes (T1D) is characterized by loss of insulin‐secreting β cells in the pancreas due to autoimmune attack by reactive lymphocytes, which can be triggered by viral infection. Proinflammatory cytokines, including interferons, are secreted by immune cells and have been implicated in pathogenesis of T1D. While interferon type II (IFNγ) has been shown to affect β cell insulin survival, its role on β cell metabolism and anti‐viral response remains to be investigated. In this study, we found that acute treatment of rat insulinoma cells (INS‐1) and Wistar rat islets with IFNγ (12 hours) induced expression of adipose triglyceride lipase (ATGL) mRNA and protein, the major enzyme that contributes to triacylglyceride (TAG) lipolysis. The increased expression of ATGL by IFNγ was associated with increased fatty acid oxidation and upregulation of genes involving in this process. Pharmacological inhibition of carnitinepalmitoyltransferase 1A (CPT1A), the rate‐limiting step of mitochondrial fatty acid oxidation, resulted in a substantial reduction in oxygen consumption rate in IFNγ‐treated INS‐1 cells. In contrast, genes that regulate lipogenesis and triglyceride synthesis were not affected by IFNγ treatment. Lipoprotein lipase (LPL) expression was suppressed, indicating a possible reduction in exogenous fatty acid delivery. These changes in lipid metabolism are concomitant with robust increase in expression of genes that involve in inflammation (STATs and NOS2), anti‐viral response (PKR, MxA), recruitment of immune cells (CXCL10) and a slight increase in endoplasmic reticulum stress markers (CHOP, ATFs). These data suggest that upon acute inflammation, pancreatic beta cell responds to IFNγ by alternating lipid metabolism flux, i.e. increasing lipolysis and fatty acid oxidation, which ultimately increase ATP production through oxidation of intracellular TAG. This change in metabolism may benefit the anti‐viral and anti‐inflammatory response of β cell in the early stage of β cell autoimmunity.

Support or Funding Information

JDRF 3‐SRA‐2014‐43‐Q‐R
Estradiol Receptor Agonists α and β Protect Against Brain Mitochondrial Dysfunction in a Model of Sleep Apnea
Authors: Laouafa Sofien , Bairam Aida , Soliz Jorge , Roussel Damien , Joseph Vincent
Published: November 2023

We tested the hypothesis that intermittent hypoxia (IH) induces a mitochondrial dysfunction in the brain of adult female rats, and that agonists of α and β estradiol receptors (ER) protect against this effect. Female rats were ovariectomized (OVX) and implanted with an osmotic pump continuously delivering vehicle (OVX‐Veh), PPT (OVX‐PPT – ERα agonist ‐ 30μg/kg/day) or DPN (OVX‐DPN ‐ ERβ agonist ‐ 100μg/kg/day). Two weeks later, the animals were exposed to intermittent hypoxia (IH‐21–10% O2 – 10 cycles/hour – 8 hours/day – 7 days) or room air (RA). We then used fresh brain cortex tissue (~2 mg) permeabilized with saponin to assess mitochondrial functions with high‐resolution respirometry (O2k‐Oroboros). With substrates of complex I (Pyruvate/Glutamate/Malate) and in presence of ADP, rats exposed to IH have a reduced O2 consumption compared to RA rats (55±7 vs 88±4 pmol O2/s/mg tissue, p<0.0001). PPT and DPN treatments prevented this alteration (O2 consumption being respectively 104±6 and 90±6 pmolO2/s/mg, p<0.0001 vs OVX‐IH rats). With the substrate of complex II (succinate) and ADP, there was no effect of IH, but O2 consumption was higher in OVX‐PPT rats compared to OVX‐IH rats. With the substrates of complexes I+II and ADP, O2 consumption was reduced by IH exposure compared to RA (51±9 vs 87±12 pmol O2/s/mg, p<0.0001), and this was partially prevented by DPN (68±10, p<0.03 vs OVX‐IH), while OVX‐PPT rats had the highest O2 consumption (125±11 pmolO2/s/mg, p<0.0001 vs OVX‐IH and OVX‐RA). H2O2 production measured by fluorometry with the substrates of complexes I+II and ADP is increased by exposure to IH compared to RA (3.2 ±0.4 vs 0.9±0.3 pmol H2O2/min/mg– p=0.002), and this is prevented by DPN and PPT (1.8 ±0.2 vs 1.6±0.2 pmol H2O2/min/mg).

We conclude that in OVX female rats, IH impairs the mitochondrial respiratory chain through the complex I leading to an increase of ROS (H2O2) production. Selective agonists of ERα and ERβ prevent the mitochondrial dysfunction. ERα appears more efficient to increase mitochondrial respiratory function in this model. This opens new perspectives for menopausal women suffering of sleep apnea.

Support or Funding Information

Founded by CIHR, Région Rhône alpes and consulat de France à Québec.
Effects of Hypoxia on Hydrogen Sulfide Production and Degradation Gene Expression Pathways
Authors: Forgan Leonard George , McNeill Bryony Alice , DeLeon Eric , Gao Yan , Olson Kenneth
Published: November 2023

Hydrogen sulfide (H2S) has emerged as an important regulator of physiological processes in multiple organ systems. One of the key functions of H2S is the regulation of vasoactivity and consequently blood‐flow. Our lab has shown potent effects of H2S and other sulfides on the vasculature of numerous vertebrate species, including vasoconstriction and vasodilation of both systemic and pulmonary arteries. Remarkably we have also shown that these effects are mimicked by hypoxia in every vertebrate class, a response which appears to be endothelium independent. Based on these observations, we hypothesised that changes in gene transcription of H2S production and degradation pathway genes by H2S and polysulfides would mimic hypoxia responses in vascular smooth muscle. In the current study we aimed to determine the effects of hypoxia and sulfides on H2S production (cse, cth, mpst, got1 and got2) and degradation (sqrdl, suox, tst, cdo1, tstd1 and ethe1) pathway genes in vascular smooth muscle using RT‐qPCR. Briefly, cultured primary bovine pulmonary artery smooth muscle cells (BPASMC) were grown until 70% confluent (passage 5) and exposed to either normoxia (21%O2/5%CO2) or hypoxia (1%O2/5%CO2) for 12 h with or without supplemental cysteine or sulfides (H2S, K2Sn, H2S2, H2S4) and the cells harvested for molecular analysis. Changes in gene expression in response to hypoxia were mimicked by H2S and polysulfides. In general, the presence of cysteine potentiated the responses to hypoxia. In the presence of cysteine, hypoxia caused a downregulation of mRNA transcript abundance of cbs and cth (P<0.001 ANOVA). In a similar way, H2S and H2S2 treatments resulted in a downregulation of cbs. In contrast, cth and mpst were unaffected by any treatment, although the intermediaries got1 and got2 were downregulated in both hypoxia and sulfide treated BPASMC. Degradation pathway genes (sqrdl, suox, tst) were downregulated in response to both hypoxia and sulfides, while cdo1 was upregulated in cysteine treatments (P<0.05 ANOVA). The housekeeping gene rn28s was not affected by any treatment. We conclude that the current data support our hypothesis that hypoxia and sulfides initiate gene transcription responses which mimic each other. Future work will include examining changes in H2S production and degradation pathway gene expression over an acute time‐course during hypoxia.
Duality of Early Life Maternal Separation Effects on Offspring Initial Development and Maternal Neuronal Activation
Authors: Fóscolo Daniela Rocha Costa , Machado Frederico Sander Mansur , Coimbra Cândido Celso
Published: November 2023

Background

Early life experiences induce behavioral and neuronal changes on most mammals, including humans. Maternal separation during the initial stages of biological development engender different modifications not only to the offspring, but also to the mother. Previously, we have shown that 2 weeks of daily episodes of maternal separation during the neonatal period, changed brain activity in response to stressful stimuli. Therefore, we investigated whether the same protocol of maternal separation evoked changes in neuronal phenotype and activity in the hypothalamus of matrices. In parallel, pups' growth and development were evaluated until adulthood.

Materials and methods

Maternal separation started at the 2nd day after delivery and consisted in taking the mother out of the cage for 180 min/day until the 14th day of life. Weaning happened on the 30th day of life and the animals were monitored until the 180th day. Matrices were euthanized on days 2, 14 and 30 in order to access expression of c‐fos in vasopressin and/or oxytocin neurons in the anterior hypothalamus. Offspring was sacrificed after 180 days of life in order to evaluate metabolic parameters such as leptin, insulin, glucose and triglycerides.

Results

Our results shown that 14 days of maternal separation modifies both the expression and c‐fos activity of oxytocin and vasopressin neurons in the PVN and SON. These modifications in the matrices persisted until the 30th day after delivery, but only in the PVN. In parallel, after 180 days of life, the offspring showed increased mesenteric fat, insulin resistance and leptin levels.

Conclusion

Our data points towards prolonged side effects of maternal separation on offspring metabolism during adulthood. Interestingly, the effect of maternal separation on the mother occurred at the hypothalamic level persisting until weaning at least. These results confirms the duality of effects elicited by maternal separation, both on the offspring and the mother.

Support or Funding Information

CAPES, CNPq, FAPEMIG and PRPq/UFMG
Structural Analysis of Inner Ear Hair Cell Mitochondria Near the Striated Organelle
Authors: Lesus Joseph , Perkins Guy , Lysakowski Anna
Published: November 2023

Mitochondria are important organelles found in every cell of the body. The current depiction of a mitochondrion, containing an inner membrane (IM), outer membrane and cristae that increase the surface area of the IM, has been the universally accepted model for decades. Although the basic function and structure of mitochondria have been studied, careful analysis of electron microscopy data has led to the identification of structurally different subpopulations of mitochondria in the inner ear (Lysakowski and Goldberg, JCN, 1997). There are several known mitochondrial‐related deafness and vestibular disorders, such as dizziness, vertigo and nystagmus (Iwasaki et al., Laryng, 2011). In order to further characterize inner ear mitochondrial subpopulations and relate them to these diseases, we used IMOD software to create 3D models of mitochondria located near different energy‐demanding cellular components. From these models, we collect quantitative data, such as crista surface area and volume and whole mitochondrion values. With these data, we can approximate a mitochondrion's energy output. The main structure of interest in this study was the striated organelle (SO), a cytoskeletal structure found in the apicolateral region of hair cells (Vranceanu et al., PNAS, 2012). We expected that mitochondria near the SO would have a fairly large energy output. It was hypothesized that there would be more points on the IM that joined with cristae membranes, the so‐called “crista junctions” (Rabl et al., JCB, 2009; Zick et al., BBA, 2009) on the side facing the striated organelle (Perkins et al., JNS, 2010). In the future, models of the different subpopulations can be compared structurally, which can lead to an explanation of differences in physiology or sensitivity to noxious agents. Exploring these differences should lead to determining the population of mitochondria affected in mitochondrial‐related deafness or vestibular disorders.

Support or Funding Information

Supported by NIH R21‐DC013181 (AL) and P41‐RR004050 (GP, ME).
Identification of the Neurotransmitter Substance P in Sural Nerve of Rats: An Immunohistochemistry Approach
Authors: Giorgetto Carolina , Simoes Ana Leda Bertoncini , Fazan Valeria Paula Sassoli
Published: November 2023

The sural nerve in rats is mainly sensory, providing innervation for the lateral side of the hind paws [1]. The substance P (SP) is a neurotensmitter known to be present in small myelinated and unmyelinated fibers, being closely related to pain tranmission to the central nervous system [2]. Thus, the presence of SP is expected in small fibers of the sural nerves. Nevertheless, morphological demonstration of the presence of SP in sural nerves of rats were not yet performed by immunohistochemestry (IHC). We developed an IHC protocol to identify the SP in sural nerve of male and female Wistar‐Kyoto rats. The animals were anesthetized and had the sural and phrenic nerves (positive control) surgically removed, immersed in Karnovsky solution (4% paraformaldehyde, 2% glutaraldehyde) and cryoprotected in solutions of sucrose with increasing concentrations. Transversal cryosections (18μm) of the nerves were incubated in rabbit polyclonal antibody anti‐substance P (1:10.000, 24h), followed by biotinylated anti‐rabbit IgG (1:500; 24h) and finally Extravidin‐HRP (1:1500, 24h). A reaction with diaminobenzidine intensified with nickel revealed immunoreactivity by light microscopy, confirming the effectiveness and viability of the IHC technique proposed by our protocol for identification of the SP in sural nerve. These results indicate the possibility of investigating neurotransmitter contents in different fiber pupulations in normal nerves and their correlation with peripheral neuropathies such as diabetes and hypertension.

Support or Funding Information

FAPESP, CNPq, CAPES and FAEPA
Antioxidant and Anti‐proliferative Activity of Fractions from Anona senegalensis Pers (Annonaceae) Stem Bark on HeLa Cells
Authors: Adisa Rahmat Adetutu , Getti Giulia T.M , Richardson Simon C.W
Published: November 2023

Anona Senegalensis Pers (Annonaceae) parts are used locally for herbal therapy, food and aesthetics. There is no scientific record of its anticancer activity on HeLa cells. The aim of the present study was to investigate the antioxidant, cytotoxic and apoptotic properties of the ethanol extracts of Anona senegalensis and its fractions on HeLa cancer cells. The HeLa cells were incubated for 24, 48 and 72 h with different concentrations of fractions of Anona senegalensis for cell viability using the MTT assay. Apoptotic cells were determined by DNA fragmentation and PE‐Annexin V assay using flow cytometry. Phytochemical screening of the ethanol extracts revealed the presence of tannin, saponin, phenol, flavonoid, alkaloid, terpenoids, reducing sugar and cardiac glycosides. Increasing concentrations of the fractions inhibited lipid peroxidation and DPPH radical scavenging activity in a dose‐dependent manner by at least 65% at 100 μg/mL. Furthermore, ethanol, hexane, aqueous, ethylacetate and chloroform fractions inhibited HeLa cells viability to different extent in a concentration‐dependent manner over the incubation periods. The IC50 values of ethanol, chloroform, hexane, ethylacetate, and aqueous fractions were 235.2, 24.3, 245, 1625.3, and 156.2μg/mL, respectively after 24h. These extract fractions caused apoptosis as shown by DNA fragmentation. Treatment of HeLa cells with the chloroform fraction induced the release of cytochrome C. The results indicate that Anona senegalensis possess strong antioxidant property and anti‐proliferative activity. The chloroform fraction is the most cytotoxic and the mechanism of anti‐proliferative activity involves apoptosis via the mitochondrial pathway through the release of cytochrome c from the mitochondria.

Support or Funding Information

Federal Republic of Nigeria Tertiary Education Fund (TETFUND) for Academic Staff Training & Development ‐ 2014
Synthesis and Antibacterial Activity of Novel Semi‐Synthetic Triterpenoids
Authors: Da Silva Gabriela Jorge , Pereira Manuela , Da Salvador Jorge
Published: November 2023

Emergence of antimicrobial resistance has become a critical public health concern worldwide. Novel antimicrobials are urgently needed, and plants with therapeutic properties constitute a rich source of biologically active compounds. Triterpenoids are a class of compounds found widely distributed in nature and have shown diverse biological activity, including anti‐tumoral activity. Structure‐activity studies have revealed that alterations in key carbons may enhance biological activity and reduce cytotoxicity. Little is known of the antibacterial activity of olealonic (OA) and ursolic (UA) acids. Moreover, the antibacterial activity of modified triterpene pentacyclic structures has yet to be characterized.

AIM

The objective of this study was to synthesize derivatives of olealonic (OA) and ursolic (UA) acids, and to test them against the antibiotic susceptible Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212 strains, and methicillin‐resistant S. aureus and vancomycin‐resistant E. faecalis of clinical origin.

METHODS and RESULTS

Initially, OA, UA, and 14 semi‐synthetic derivatives were screened for antibacterial activity by the disk diffusion method. Minimum inhibitory concentrations (MIC) were determined by microdilution method. MICs ranged from 62.5 to 125 ug/mL in Gram negative bacteria, from 31.3 to 125 ug/mL for S. aureus strains and from 7.8 to 125 ug/mL for E. faecalis. In general, UA and compound 14, a derivative of UA, showed the best activity against Gram positive bacteria. No significant differences in MICs were observed between the antibiotic susceptible and resistant clinical strains. Both compounds reduced the bacterial growth of S. aureus ATCC 29213 and E. faecalis ATCC 29212 in the presence of 0,5xMIC, MIC, 2xMIC relatively to the control, more significant in the latter. MTT cytotoxicity assays revealed that compound 14 did not induce significant cell toxicity in macrophages (RAW 264.7), with 84% viability at concentrations delivering effective (MIC). Overall, the compounds tested showed a better antimicrobial activity against Gram‐positive bacteria.

CONCLUSION

The data indicate that compound 14 may offer a new avenue for the development of new antimicrobials with reduced cytotoxicity. This work also illustrates that modifications of the natural molecule and the study of structure‐activity relationship is crucial to unravel new biological activity.