bluuebunny/crossref_metadata_2025 huggingface dataset:
Sodium content in US packaged foods, 2009
Authors: Gillespie Cathleen , Maalouf J. , Yuan K. , Cogswell M. E. , Gunn J. , Levings J. , Moshfegh A. , Merritt R.
Published: June 2021

In 2010, the Institute of Medicine recommended gradual reductions in the sodium content of packaged and restaurant foods as a primary strategy to reduce US sodium intake. With increased focus and voluntary efforts by some manufacturers to reduce the sodium in their products, monitoring sodium content of packaged foods overtime will aid the evaluation of progress. Using commercially available data, we created a database with sales and Nutrition Facts Panel information for over 8,000 packaged food products sold in US grocery stores in 2009. Using this database, we estimate the sales‐weighted mean and distribution of sodium concentration of packaged foods within food groups that contribute the most dietary sodium to the US diet including the food categories that comprised the top contributors to dietary sodium intake in 2007–2008. We also estimated the proportion of products within each category that exceed 480 or 600 mg, the Food and Drug Administration's limits for the sodium content in “healthy” foods and meals, respectively. Products in the pasta mixed dishes category had the highest mean sodium content per serving at 966 milligrams (mg), and the sodium content per serving in this food group ranged from a minimum of 80 mg to a maximum of 1,960 mg. The interquartile range for all categories was 90 to 500 mg. More than 75% of the products sold in 7 of the 20 food categories examined exceeded 480 mg/serving. In 3 of these categories, pizza, mixed meat dishes, and pasta mixed dishes, over 75% of the products sold exceeded 600 mg/serving. Given the percent of products exceeding limits for healthy foods and the range of sodium content in products in the top food categories contributing to US sodium intake, there is potential for sodium reduction.
Iron Biofortified Pearl Millet Improves Iron Status in Indian School Children: Results of a Feeding Trial
Authors: Haas Jere D. , Finkelstein Julia L. , Udipi Shobha A , Ghugre Padmini , Mehta Saurabh
Published: June 2021

Objective

To examine the effects of an iron biofortified pearl millet intervention on iron status in children.

Methods

246 children (11–18 y) were enrolled in a randomized trial of an iron biofortified pearl millet intervention (vs. control pearl millet), daily for a 6 month period in India. Iron status (hemoglobin, ferritin, sTfR), inflammation (CRP, AGP), and anthropometric indices were determined at enrolment, 3, and 6 months. Linear and binomial regression models were used to evaluate the effect of the intervention on iron status in children.

Results

43.6% of children were iron deficient (serum ferritin <15.0 μg/L) and 27.9% were anemic (Hb <12.0 g/dL) at baseline. After 3 months there was an overall effect of the iron biofortified pearl millet intervention on serum ferritin in previously iron deficient subjects. Also, the intervention resolved 65% of iron deficiency in children, and was associated with a two‐fold increase in the likelihood of resolving iron deficiency at 6 months (RR: 1.94, 95% CI: 1.15–3.27; P=0.01).

Conclusions

Supplementation with iron biofortified pearl millet significantly resolved iron deficiency in children within 6 months.

Funding Source: Harvest Plus
A diet high in saturated fat and low in calcium is more detrimental to bone health of male Wistar rats than a fructose‐rich diet
Authors: Castelli Laura M , Miotto Paula M , Bruce W Robert , Ward Wendy E
Published: June 2021

Dietary factors can have major metabolic effects through a pathway to energy excess, insulin resistance, insulinemia and diabetes; or through a pathway to decreased calcium availability, intestinal barrier failure, bacterial translocation, inflammation and oxidative stress. The latter path is known to affect bone structure and strength, but there is no direct evidence on a possible interaction with processes affecting the development of insulin resistance. Our objective was to determine which dietary components – high saturated fat and low calcium (referred to as the oxidative stress diet) or high fructose ‐ is more detrimental to bone strength and quality in male Wistar rats. Rats were fed 1 of 4 diets for 8 weeks: control, oxidative stress diet (OX), oxidative stress plus fructose diet (OX + fructose), or fructose diet. Strength at femur midpoint and neck, and third lumbar vertebra, were measured using a materials testing system. Whole femur, 1/3 proximal femur, and lumbar spine bone mineral content (BMC) and bone mineral density (BMD) were measured using dual x‐ray absorptiometry. Body weights of fructose‐fed rats were lower (P=0.016) than OX‐fed rats. Rats fed the OX and OX + fructose diet had lower (P<0.001) peak load, BMC and BMD compared to rats fed control and fructose diet. Diets rich in saturated fat and low in calcium resulted in weaker femur and spine, but a diet rich in fructose did not affect bone health.
Effects of iron deficiency on the production of proinflammatory cytokines in 8 to 12 year‐old children
Authors: García‐Miranda Luis Armando , Estrada José Antonio , Paredes‐Cervantes Vladimir , Contreras Irazú
Published: June 2021

Iron is an essential nutrient for many metabolic processes, which include erythropoiesis, oxidative metabolism and the immune response. Iron deficiency (ID) is considered a worldwide public health problem. The objective of this study was to determine the prevalence of ID and its effects on the production of proinflammatory cytokines (TNF‐α and INF‐γ) in 8 to 12 year‐old children from a community in the State of Mexico. Blood samples were obtained from 100 children; weight and height were also measured. Serum was used to determine iron concentrations by spectrophotometry, as well as TNF‐α and INF‐γ production by ELISA. In addition, a complete blood count was performed. ID was considered with values ≤ 60 μg/dL. We observed a high prevalence (>;50%) of overweight children; ID was determined in 19% of children. The percentage of children with iron deficiency anemia (Hb ≤ 11 g/dl) was 5%. Blood differential showed a decreased frequency of lymphocytes and increased frequency of neutrophils in children with ID, although the total number of leucocytes was unaltered. INF‐γ production was reduced 16% in the ID group compared to the control group. We observed no difference in TNF‐α production between groups.
Vitamin D regulates cross talk between epithelial cells and immune cells through the secretion of CD14
Authors: Simmons Katrina , Welsh JoEllen
Published: June 2021

Inflammatory signaling between epithelial cells and immune cells contributes to a variety of diseases including cancer, autoimmune disorders, and obesity. Vitamin D, a potent immunomodulator, has been shown to regulate immune responses and reduce the risk of infections by acting on immune cells, yet its effects on the immune functions of epithelial cells and their impact on the immune microenvironment in epithelial tissues are less clear. We have demonstrated that 1α,25‐dihydroxyvitamin D (1,25D) dramatically increases the expression of the pattern recognition receptor CD14, which binds microbial lipopolysaccahride (LPS), in human mammary epithelial (HME) cells. We found through ELISA and microscopy that 1,25D enhances the accumulation of soluble not membrane‐localized CD14 and that conditioned media from 1,25D‐treated HME cells, which contains soluble CD14, promotes anti‐inflammatory responses from macrophages when exposed to LPS. Studies with the vitamin D receptor (VDR) analog JN, a non‐genomic VDR agonist, found JN induces CD14 expression and secretion comparable to 1,25D suggesting the membrane‐localized VDR contributes to 1,25D's induction of CD14. Overall this study supports the concept that 1,25D stimulates the secretion of CD14 from epithelial cells through the VDR to promote an anti‐inflammatory environment.

Grant Funding Source: National Center for Complementary and Alternative Medicine Predoctoral F31 NRSA Fellowship
Dynamic Model Predicting Overweight, Obesity, and Extreme Obesity Prevalence Trends
Authors: Thomas Diana Maria , Weederman Marion , Fuemmeler Bernard , Martin Corby , Dhurandhar Nikhil , Bredlau Carl , Heymsfield Steven B. , Ravussin Eric , Bouchard Claude
Published: June 2021

Objective

To model known multiple population parameters associated with changes in the prevalence of body mass index (BMI) classes (overweight, obese, and extremely obese) and to establish the necessary conditions under which the prevalence of obesity will eventually plateau.

Methods

A six differential equation system was developed that predicts population‐wide obesity prevalence trends. The model considers both social and non‐social influences on weight gain, incorporates other known parameters affecting obesity trends, and allows for country specific population growth. Long‐term obesity trends were calculated after input of parameters specific to the US.

Results

The dynamic model predicts that the prevalence at which obesity plateaus in a population is primarily a function of birth rate and the probability of being born in an obesogenic environment; that obesity prevalence will eventually plateau independent of current prevention strategies if key population‐specific parameters such as birth rate remain constant; and that the US prevalence of obesity, overweight, and extreme obesity will plateau by about 2030 at 28%, 32%, and 9%, respectively.

Conclusions

The US prevalence of obesity is tapering off and will plateau in less than two decades, independent of current preventative strategies. This trend has important implications in accurately evaluating the impact of various anti‐obesity strategies aimed at reducing the national obesity prevalence.
Gender‐specific associations of socioeconomic factors and obesity among urban adults in Turkey
Authors: Fitzgerald Nurgul , Turnagol Husrev , Basoglu Sevil , Kosar Nazan
Published: June 2021

This study examined the associations between socioeconomic factors and obesity in a nationally representative sample of adults (≥19) in 13 urban centers in Turkey. Data were collected through face‐to‐face household interviews; anthropometric measurements were conducted by study assistants. The analytical sample included 4,813 adults with measured weight, height, and weight circumference (WC) values. Logistic regression analyses were used to analyze the determinants of obesity (BMI≥30.0) and high WC (≥102 cm for men, ≥88cm for women).

The average age in the sample was 41.7yr, 61.5% were females, and 58.3% reported eighth grade or less education. Obesity and high WC were more common among women than men (34.3 vs 14.9% and 51.0 vs 15.8%, respectively, p<0.001). After adjusting for age, migration, home ownership and health insurance status, obesity was linked to being married, having a car and less than college education among women; education, migration, and being married were also positively related to having a high WC. Among men, not having health insurance was linked to less likelihood of obesity and high WC (p<0.05).

Relatively high rates of overall and central obesity among adults in urban Turkey seem to have gender‐specific associations with individual and household level socioeconomic characteristics.
Lower levels of leptin and adiponectin independent of body mass index in Japanese American women: The Multiethnic Cohort
Authors: Maskarinec Gertraud , Morimoto Yukiko , Kim Yeonju , Lim Unhee , Cooney Robert V , Conroy Shannon M , Franke Adrian A , Wilkens Lynne R , Hernandez Brenda Y , Goodman Marc T , Le Marchand Loic , Kolonel Laurence N
Published: June 2021

Ethnic differences in obesity and body fat distribution may contribute to varying chronic disease risks, e.g., for breast cancer and diabetes. To explore if adipokines mediate this connection, we evaluated the relation of body mass index (BMI as defined by WHO) and serum levels of leptin and adiponectin measured by ELISA among 936 white, Japanese American (JA), African American (AA), Latino (LA), and Native Hawaiian (NH) female control participants within the Multiethnic Cohort. BMI differed significantly by ethnicity (p<0.0001). As compared to whites (25.3±5.2 kg/m2), BMI was lower in JA (23.7±3.8 kg/m2) and higher among AA, LA, and NH (28.9±5.8, 28.0±5.2, 28.3±5.9 kg/m2, respectively). Linear models were applied to compare means of log‐transformed biomarkers by adiposity status and ethnicity. In all ethnic groups, leptin was higher in overweight and obese than in normal weight women, whereas adiponectin showed an inverse trend (p<0.0001 for all). JA women had significantly lower leptin and adiponectin levels than whites across the 3 BMI categories; the respective differences between the two ethnic groups were 4.0, 8.7, and 18.9 ng/mL for leptin (p=0.0004) and 5.9, 4.4, and 3.6 μg/mL for adiponectin (p<0.0001). The higher obesity‐related disease risk in JA and other ethnic groups may be in part mediated by differences in adipokine either stemming from endocrine genetics or fat distribution.
Influence of long‐term exercise training followed by a diet/exercise weight‐loss intervention on body composition and metabolic variables in subjects with pre‐diabetes
Authors: Starr Kathryn N , Granville Esther O , Ocampo Christine I , Slentz Cris A , Bateman Lori A , Willis Leslie H , Rose Sarah B , Kraus William E , Bales Connie W
Published: June 2021

Lifestyle measures such as diet and exercise are strongly encouraged to reduce diabetes risk; however, studies of longer than 6 month's duration are rare and strategies for long‐term adherence remain elusive. The goal of this study was to examine the health impact of a 12‐month lifestyle intervention in women and men (n =21; mean age = 61.2 yrs) who had pre‐diabetes at baseline, defined as a fasting blood glucose (FBG) of 95–124 mg/dl on 2 occasions. Body mass (BM) and composition, namely fat (FM in kg and %) and fat‐free (FFM in kg and %) mass by BodPod, waist circumference (WC), hip circumference (HC) and plasma levels of total (TC) and HDL‐cholesterol (HDL‐C), triglycerides (TG) and FBG were assessed at baseline (B), after 6 months of aerobic exercise training (6‐ET), and after an additional 6 months of combined diet/aerobic exercise (12‐ET). Compared to B, values at 6‐ET were lower (p<0.001) for BM, FM %, FM kg, WC, and HC, and higher for FFM % and HDL‐C. At 12‐ET compared to 6‐ET, there were significant improvements in BM (p<0.0001), FM % (p<0.05), FFM kg (p<0.01), FFM % (p<0.05), WC (p<0.0001), HC (p<0.001), TG (p<0.01), and HDL‐C (p<0.01). FBG, LDL‐C, and TC were essentially unchanged during the trial. Our findings show that adding a weight loss and exercise treatment after 6 months of aerobic exercise confers additive improvements in body mass and composition and in TG and HDL‐C. Supported by NIDDK R01‐DK081559 and NIA AG000029.
Serum and Dietary Advanced Glycation End Products in Patients with Diabetic Foot Ulcers
Authors: Maier Haiyan Min , Spicer Maria T. , Arjmandi Bahram H.
Published: June 2021

Diabetic foot ulcers (DFU) occur in patients with a history of poorly controlled blood glucose. Advanced glycation end products (AGEs) may influence the occurrence of DFU due to elevated oxidative stress (OS) and inflammation. This study examined the association between AGEs and serum markers of OS and inflammation in diabetic patients with (DFU) and without diabetic foot ulcer (DM).

Eighty adult participants were recruited and categorized into one of three groups: non‐diabetic control (C), DM, and DFU. Twenty‐four hour food recalls, foot ulcer history and blood samples were collected from each participant for analyses. Dietary and serum AGEs, tumor necrosis factor‐α (TNF‐α), and thiobarbituric acid reactive substances (TBARS) were determined. Using SPSS 21, one way ANOVA and Pearson correlation were used for statistical analyses.

The diabetic groups had significantly higher levels of dietary AGEs and TNF‐α than C (P < 0.05). Serum AGEs and TBARS were elevated in DFU, compared with C (P < 0.05). TBARS were significantly higher in DFU than DM and C (P < 0.5). Serum AGEs and TBARS were significantly correlated with dietary AGEs (P < 0.01).

Both dietary and serum AGEs were the highest in DFU. Dietary AGEs modulates serum AGEs concentration. Reducing dietary AGEs intake may benefit patients with DFU by decreasing OS.

Grant Funding Source: Florida State University
Determinants of iron‐deficient anemia among food insecure Brazilian infants
Authors: Lemos Thaisa , Vitolo Marcia Regina , Hoffman Daniel J.
Published: June 2021

Iron‐deficient anemia (IDA) is still considered a public health problem in developing countries. Brazil's latest national data estimated that IDA affected 21% of children under 5 years of age. We investigated whether the prevalence of IDA (defined as hemoglobin < 11g/dL) and its determinants would differ between food secure (FS) and insecure (FI) infants. From the 284 infants, 53.5% were FS and 61.6% had IDA. There were no statistical differences in prevalence of IDA between FI and FS infants, although FI infants had lower mean Hb level. After controlling for confounders, maternal age (odds ratio [OR] = .917, 95% Confidence Interval [CI] = .857‐.981) and BMI (OR = .384, 95%CI = .166‐.888) were negatively associated with IDA at 12 months, while household size (OR = 1.481, 95%CI = 1.040–2.109) and introduction of soda before 10 months (OR = 2.377, 95%CI = 1.041–5.428) were positive predictors for FS infants. For FI children, only the cluster “good practices” (solid food after 4 months, breastfeeding for 6+ months, lipid and sugar dense food after 12 months) remained significant (OR = 2.68, 95%CI = 1.061–6.772). In conclusion, among low‐income FS or FI infants, there is no statistical difference in the prevalence of IDA, although determinants are different between the two food security levels. Funding was provided by the Brazil National Funding for Research and Capes Foundation of the Brazilian Ministry of Education.
Effects of folylpolyglutamate synthase modulation on CpG promoter DNA methylation and gene expression in human colon cancer cells
Authors: Kim Sung‐Eun , Hinoue Toshinori , Kim Michael , Sohn Kyoung‐Jin , Weisenberger Daniel , Laird Peter W. , Kim Young‐In
Published: June 2021

Folylpolyglutamate synthase (FPGS) facilitates intracellular retention of folate by polyglutamylation, thereby playing a critical role in folate homeostasis. We investigated whether FPGS modulation would affect CpG promoter methylation and gene expression profiles in human HCT116 colon cancer cells. Illumina HT‐12 and Infinium Methylation assay were used, and functional analysis was performed by Ingenuity Pathway Analysis. We validated mRNA expression of selected genes by qRT‐PCR. 2897 genes (most involved pathways: cell cycle and DNA replication, recombination, and repair) were differentially expressed in the FPGS‐overexpressed cells, while 359 genes (most involved pathways: cell death and cell cycle) were differentially expressed in the FPGS‐inhibited cells. 864 and 626 genes showed altered CpG promoter methylation associated with FPGS overexpression and inhibition, respectively. An integrated analysis revealed 65 and 6 differentially expressed genes involved in cell cycle, cell death, and cell‐to‐cell signaling and interaction, expression of which was regulated by CpG promoter methylation changes associated with FPGS overexpression and inhibition, respectively. FPGS modulation can affect differential gene expression and CpG promoter methylation involved in important biological pathways, and some of the observed differential gene expression appear to be epigenetically regulated.

Grant Funding Source: CIHR Fund 14126
Characterization of a GCN2/eIF2alpha independent pathway that regulates the expression of eukaryotic initiation factor 4E‐binding protein 1
Authors: Mazor Kevin Michael , Stipanuk Martha
Published: June 2021

Lack of essential amino acids is known to activate the eIF2α/ATF4 stress signaling pathway via activation of GCN2 (eIF2α kinase 4) by nonaminoacylated tRNAs. In previous studies, we have observed that eukaryotic initiation factor 4E‐binding protein 1 (4EBP1) expression is upregulated in methionine (Met)‐deficient cells even in the absence of an active GCN2/eIF2α signaling pathway. To further characterize this GCN2/eIF2α‐independent pathway, we compared the effects of Met and histidine (His) deficiency in murine embryonic fibroblasts (MEFs). In wild‐type MEFs, either –His or –Met medium resulted in increases in 4EBP1 mRNA and protein abundance as well as ATF4 mRNA levels. This upregulation of 4EBP1 and ATF4 was also seen in MEFs lacking GCN2 or with mutant eIF2α when they were cultured in –Met medium, but not when cultured in –His medium. These results imply the existence of a GCN2‐and eIF2α phosphorylationin‐dependent pathway that is capable of increasing the expression of both 4EBP1 and ATF4 in response to Met but not His deficiency. Supported by Grants DK‐083473 and T32DK007158 (NIDDK) and by the Division of Nutritional Sciences.
Lipolytic effect of olive leaf extracts and a major component on mature 3T3‐L1 cells
Authors: Ryu Su‐Jung , Choi Hyeon‐Son , Lee Ok‐Hwan , Lee Boo‐Yong
Published: June 2021

The olive, Olea europaea, is a small tree native to the countries adjoining to areas of eastern Mediterranean basin and southern Caspian Sea. Recent studies showed biological activities such as antioxidative, hypoglycemic, antihypertensive, antiatherosclerotic, and antiadipogenic effects from olive leaves. In this study, we evaluated the effect of olive leaf extract (OLE) on lipolysis in mature 3T3‐L1 cells by examining the levels of free glycerol, triacylglycerol, and lipolytic enzymes. OLE increased the release of free glycerol from triacyglycerol in a dose‐dependent manner. The mRNA expression levels of hormonal sensitive lipase (HSL), adipose triglyceride lipase (ATGL) and perilipin showed no significant differences in the presence and absence of OLE. The protein levels of the major lipolytic enzymes, HSL and ATGL, were not affected by OLE treatment. However, OLE increased the level of phosphorylated HSL, an indicator of lipolytic activation, which led to a decrease in the triacylglycerol level. In addition, oleuropein, a major compound in OLE, decreased the level of triacylglycerol and increased the level of free glycerol, like OLE. The protein levels of HSL, p‐HSL, and ATGL are increased by oleuropein treatment. Our data suggest that oleuropein in OLE stimulates lipolysis through activation of HSL in adipocytes.
Dual activation of p38MAPK and SFK pathways is required to induce endothelial permeability
Authors: Adam Alejandro Pablo , Lowery Anthony , Vincent Peter
Published: June 2021

We previously showed that activation of endogenous Src family kinases (SFK) by expression of a dominant negative form of Csk (DN‐Csk) is not sufficient to induce changes in endothelial monolayer permeability, but allows low doses of TNF to increase endothelial permeability. This increase induced by joint SFK activation and TNF treatment is attenuated by addition of p38 inhibitors. Thus, we hypothesized that p38MAPK activation may induce an increase in permeability of ECs already primed by activation of SFKs. To test this, we constructed a lentiviral vector to express a flag tagged constitutively active form of MKK6 under the control of a tetracycline‐regulated promoter (iMKK6E). Addition of doxycyclin (dox) to infected cells induced a dose‐dependent increase in MKK6E expression resulting in the phosphorylation and nuclear translocation of p38 as well as HSP27 phosphorylation. This treatment did not induce any change in permeability. In contrast, MKK6E expression resulted in a large drop in TEER and an increase in albumin clearance in cells with SFK activation. This was accompanied by the formation of large gaps in the monolayer but no change in actin fibers. Our results support a model in which dual activation of SFKs and p38 are required for the induction of endothelial permeability. This model could explain the tight spatio‐temporal response to inflammatory mediators during an inflammatory response.
BNIP3 regulates AT101 induced cytotoxicity in MPNST cells
Authors: Kaza Niroop , Carroll Steven L , Roth Kevin A
Published: June 2021

Malignant peripheral nerve sheath tumors (MPNSTs) are highly aggressive Schwann cell‐derived sarcomas and a leading cause of mortality in neurofibromatosis type 1 (NF1) patients. Current treatment modalities have been ineffective resulting in a high rate of recurrence and poor five year survival, which necessitates the exploration of novel therapeutic agents. AT101, a BH3 mimetic, has been shown to induce cell death in several cancers and is in phase II clinical trials. The objective of this study was to investigate the molecular mechanisms regulating AT101 induced cytotoxicity in human MPNST cells and to evaluate its chemotherapeutic potential for MPNSTs. AT101 caused a concentration dependent decrease in viability in MPNST cells which was not accompanied by caspase activity. AT101 also induced an increase in autophagic flux and inhibiting induction of autophagy attenuated cell death induced by AT101. Additionally, AT101 caused increased expression of BNIP3, an atypical BH3‐ only protein involved in cell death, along with increased levels of HIF‐1α protein, a transcriptional regulator of BNIP3, in MPNST cells. Moreover, siRNA mediated knockdown of BNIP3 significantly attenuated AT101 induced cell death. In conclusion, AT101 induces an increase in BNIP3 expression and autophagy leading to cell death independent of caspase activation in MPNST cells. This work was supported by grants from NCI (RO1 CA134773).
Matrix metalloproteinase‐28 deletion attenuates early cardiac dysfunction following myocardial infarction by restraining neutrophil infiltration and limiting the inflammatory response
Authors: Ma Yonggang , Yabluchanskiy Andriy , Zhang Jianhua , Ramirez Trevi A. , Manicone Anne M. , Lindsey Merry L.
Published: June 2021

By 1 week post‐myocardial infarction (MI), MMP‐28 null mice show exacerbated cardiac rupture and dysfunction associated with inhibition of M2 macrophage activation, extracellular matrix deposition, and collagen cross‐linking. The impact of MMP‐28 deletion on the early inflammatory response, however, remains to be evaluated. C57BL/6J wild type (WT, n=12) and MMP‐28 null (n=12) mice at 3–6 months of age were subjected to permanent coronary artery ligation and sacrificed at 1 day (d). Ejection fraction declined from 62 ±1% at baseline to 17±1% at d1 in WT (p<0.05), and this decline was attenuated 24% by MMP‐28 deletion (p<0.05). In the infarct region, 21 out of 84 inflammatory genes increased and 16 decreased in WT, while 21 were elevated and 11 reduced in MMP‐28 null mice, compared to corresponding d0 values (all p<0.05). Ccl22, Ccr8, Cxcr5, Il13, Il1α, Il1f6, and Ltb were lower in MMP‐28 null (all p<0.05), suggesting reduced recruitment or activation of neutrophils. By immunohistochemistry, 28% fewer neutrophil infiltration was noted in MMP‐28 null. In conclusion, MMP‐28 deletion attenuated d1 cardiac function by inhibiting neutrophil infiltration to limit the early inflammatory response.
Bile acid depletion increases susceptibility to acetaminophen‐induced hepatotoxicity in mice
Authors: Bhushan Bharat , Borude Prachi , Edwards Genea , Walesky Chad , Cleveland Joshua , Li Feng , Ma Xiaochao , Apte Udayan
Published: June 2021

Bile acids are known to play significant role in liver homeostasis. However, the role of bile acids in pathogenesis of acetaminophen (APAP)‐induced acute liver failure (ALF) is not known. Here we report that depletion of bile acids using cholestyramine (CSA), a bile acid sequestering resin, increases susceptibility of C57BL6 mice to APAP‐induced hepatotoxicity. C57BL6 mice were fed either normal diet or 2% CSA containing diet for 1 week, treated with 400 mg/kg APAP (ip) and analyzed over a time course of 0 to 24 hr. The CSA‐fed mice developed rapid and higher liver injury after APAP treatment. Depletion of bile acids by CSA did not alter metabolism of APAP. However, CSA‐fed mice exhibited rapid and significantly higher activation of c‐Jun N‐terminal protein Kinase (JNK) following APAP treatment. Furthermore, a significant decrease in intestinal fibroblast growth factor 15 (FGF15) mRNA was observed in CSA fed mice before and after APAP treatment. Compensatory liver regeneration following APAP overdose was delayed in the CSA‐fed mice up to 12 hr but was not different than the normal diet fed mice at 24 hr after APAP treatment. Taken together, these data indicate that depletion of intestinal bile acid increases susceptibility to APAP by rapid and higher hepatic JNK activation. These data indicate that bile acids and FGF15‐mediated gut‐liver signaling axis play an important role in cytoprotection following APAP‐induced ALF.

These studies were supported by NIH ‐ P20 RR021940 (Udayan Apte), and AASLD/ALF Liver Scholar Award (Udayan Apte)
Coping with Unexpected Environments – Implications for Epigenesis
Authors: Soen Yoav
Published: June 2021

Developing organisms have evolved a wide range of mechanisms for coping with recurrent environmental conditions. How they cope with rare or unforeseen challenges is, however, unclear as are the implications to subsequent generations of non‐exposed offspring.

We investigate these questions using a synthetic drug/anti‐drug system which allows us to confront the development of the fly, D. melanogaster, with artificial distributions of toxic stress that are not expected to occur during fly development. Survival of the flies in this system depends of their ability to modify their development. We found that under a wide range of toxic scenarios, the challenge modifies the otherwise robust patterns of development, resulting in changes in gene expression as well as in the rate of larval development and adult morphology (in some of the cases). We show that part of this response is enabled by suppression of Polycomb group genes (PcG), which leads to de‐repression of developmental regulators and their expression in new domains, hence the change in developmental patterns. Remarkably, some of the developmental alterations were non‐genetically inherited by subsequent generations of unchallenged offspring suggesting that the challenge also modifies the germline of the flies.

This was indeed confirmed by analysis of maternal RNA in eggs of challenged versus unchallenged flies. These results reveal a process of epigenesis by which stressful, non‐familiar environment suppresses the Polycomb system and induces developmental modifications that persist across generations through non‐Mendelian mechanisms.
Human and bovine osteopontin from milk and recombinant human osteopontin may stimulate intestinal proliferation and immune functions via various mechanisms revealed by microarray analysis
Authors: Jiang Rulan , Lonnerdal Bo
Published: June 2021

Osteopontin (OPN) is a highly acidic phosphorylated glycoprotein, present at high concentration in human milk and low concentration in bovine milk and infant formula. Human OPN (hOPN) and bovine OPN (bOPN) have been demonstrated to be resistant to in vitro digestion. Thus, hOPN and bOPN may play important roles in intestinal and immunological development in infancy. hOPN, recombinant hOPN (rhOPN) and bOPN may function differently due to different post‐translational modifications and 63% amino acid identity between hOPN and bOPN. Therefore, we hypothesized that hOPN, rhOPN and bOPN are capable to stimulate intestinal proliferation and regulate immune functions via various mechanisms. BrdU assays showed that hOPN, rhOPN and bOPN significantly stimulated proliferation of human intestinal cells (HIEC). Compared with hOPN and rhOPN, a lower concentration was needed for proliferation effects exerted by bOPN. To screen for underlying mechanisms by which hOPN, rhOPN and bOPN stimulate intestinal proliferation, microarray assays (Illumina) were performed. Selected genes were verified by RT‐PCR with a high validation rate. Genes significantly regulated by hOPN, rhOPN, and bOPN were 243, 356, and 284 respectively of which only 50 genes were modulated by all. Genes tightly related to proliferation and immune function were significantly modified by hOPN, eg MAPK13, CCNE1, CdGAP, CXCL10, IL6ST and NFKB. Some of these genes were affected by rhOPN or bOPN, but not all. In summary, hOPN, rhOPN and bOPN are capable to stimulate intestinal proliferation and regulate immune functions but via various mechanisms.
Structural Insights into O‐GlcNAc Transferase
Authors: Walker Suzanne
Published: June 2021

O‐GlcNAc Transferase (OGT) is an essential glycosyltransferase that catalyzes the attachment of N‐acetylglucosamine (GlcNAc) to serines and threonines of numerous nuclear and cytoplasmic proteins. Global O‐GlcNAcylation levels increase with increased flux through the hexosamine biosynthetic pathway, which produces OGT's donor substrate, UDP‐GlcNAc. Increased O‐GlcNAcylation is associated with widespread changes in gene expression, but the molecular mechanisms underlying these changes remain poorly understood. It was recently shown that OGT catalyzes another post‐translational modification: it promotes the cleavage of host cell factor 1 (HCF‐1), a ubiquitous transcriptional coregulator found in several chromatin remodeling complexes. We will describe mechanistic studies of OGTmediated cleavage of HCF‐1 and will present structures of OGT bound to HCF‐1 proteolytic repeats. Implications of these studies are numerous.
From force‐sensing to power generation – How force modulates the ATPase kinetics of myosin motors
Authors: Ostap E Michael , Greenberg Michael J , Laakso Joseph M , Goldman Yale E , Shuman Henry
Published: June 2021

Myosins are motor proteins that hydrolyze ATP to power various cellular processes including muscle contraction, intracellular cargo transport, cytoskeletal anchoring, and signaling. Most members of the myosin family follow the same kinetic pathway for ATP hydrolysis, where force‐generating structural changes are linked to the release of inorganic phosphate and ADP. Interestingly, mechanical loads on myosins modulate the rate constants of key steps on the ATPase pathway, resulting in load‐dependent changes in velocity and power production. We investigated the load sensitivity of two members of the class‐1 family of myosins (Myo1b and Myo1c) using single molecule optical‐trap assays and ensemble biochemical techniques. We determined that the unloaded ATPase and transient kinetic rate constants of these two myosins are similar. Surprisingly, we found the kinetics of Myo1b to be substantially more sensitive to mechanical load than Myo1c. We also determined that load affects different biochemical steps in these two myosins, despite their having nearly identical unloaded kinetic properties. Understanding the different load sensitivities of these motors provides insights into their respective cellular roles (Myo1b is a tension‐sensing anchor and Myo1c is a power‐generating transporter) as well as the mechanisms of force sensing within the larger myosin family. Supported by NIH grant, PO1 GM087253.
Substituting dietary monounsaturated fat for saturated fat lowers inflammasome activation observed on a high saturated fat diet
Authors: Kien Craig Lawrence , Crain Karen I. , Bunn Janice Y. , Fukagawa Naomi K. , Poynter Matthew E.
Published: June 2021

Altered lipid homeostasis (obesity) is linked to inflammation and defective insulin signaling. In the presence of palmitic acid (PA), but not oleic acid (OA), lipopolysaccharide (LPS)‐stimulated macrophages activate the NLRP3 inflammasome, secreting augmented levels of Interleukin (IL)‐1β and IL‐18 (Nat Immunol12:408). We hypothesized that lowering the PA/OA ratio of the PA‐rich, Western Diet would alter cytokine secretion from peripheral blood mononuclear cells (PBMC) harvested from 14 fasted, young adults (8 men/6 women), enrolled in a randomized, double‐masked, cross‐over trial comparing two 3‐week diets: (1) high PA (HPA), with a fatty acid pattern resembling their habitual diet; (2) low PA/high OA (HOA). Immediately after collection, PBMC were stimulated with 1 ng/ml LPS for 24 hr, and cytokines were measured. Log (base 10) transformed data, corrected for baseline values on a low fat diet, revealed that IL‐1β (P = 0.029), IL‐18 (P = 0.037), IL‐10 (P = 0.02), and Tumor Necrosis Factor‐1α, (P = 0.039) were higher during HPA, compared to HOA. These data suggest that, in healthy adults, the higher than recommended PA intake of the Western Diet facilitates augmented secretion of several cytokines, including those regulated by inflammasome activation. (Support: NIDDK: R01‐DK 082803).
INTEGRATIVE GENOMIC STRATEGIES FOR BIOMARKER DISCOVERY IN BREAST CANCER IN MINORITY POPULATIONS
Authors: Hicks Chindo
Published: June 2021

Breast cancer remains one of the leading causes of death among women in the United States. It is the most commonly diagnosed cancer among African American women with an estimated 20,000 new cases and 6,000 deaths each year. Currently, African American women have a 27% higher death rate than white women. Therefore, there is an urgent need to identify molecular markers to eliminate health disparities in breast cancer. Advances in high‐throughput microarray, genotyping and more recently next generation sequencing technologies have made possible identification of molecular signatures, genetic variants and driver mutations associated with breast cancer. However, there has been a strong bias towards use of samples from populations of European ancestry in biomarker discovery in breast cancer. To date, the massive amounts of data generated by microarray, genotyping and sequencing technologies have not been maximally leveraged and integrated to identify biomarkers associated with breast cancers in minority populations. We shall present novel integrative genomic strategies which integrate gene expression, genotype and sequence data to identify molecular markers and biological pathways involved in breast cancer in minority populations. We have chosen as our model the most aggressive subset of breast cancer, the triple negative breast cancer (TNBC), which disproportionately affects African American women.
Fluid Shear Stress induces the Clustering of Heparan Sulfate via Mobility of Glypican‐1 in Lipid Rafts
Authors: Zeng Ye , Ebong Eno , Rizzo Victor , Tarbell John
Published: June 2021

The endothelial glycocalyx plays important roles in mechanotransduction. We recently investigated the distribution and interaction of glycocalyx components on statically cultured endothelial cells. In the present study, we further explored the unknown organization of the glycocalyx during early exposure (first 30 min) to shear stress using confocal microscopy and tested the hypothesis that proteoglycans with glycosaminoglycans, which are localized in different lipid microdomains, respond distinctly to shear stress. During the initial 30 minutes, we observed that heparan sulfate (HS) and glypican‐1 clustered in the cell junctions. In contrast, chondroitin sulfate, bound albumin and syndecan‐1 did not move. The caveolae marker, caveolin‐1, did not move indicating that caveolae are anchored sufficiently to resist shear stress. Shear stress induced significant changes in the distribution of ganglioside GM1 (a marker for membrane rafts labeled with cholera toxin B subunit – CTx‐B). Thus, fluid shear stress induced the cell junctional clustering of lipid rafts with their anchored glypican‐1 and associated HS. In contrast, the mobility of chondroitin sulfate, transmembrane bound syndecan‐1, and caveolae were constrained during exposure to shear stress. This study illuminates the role of changes in glycocalyx organization that underlie mechanisms of mechanotransduction.

Grants: Supported by NIH Grant HL 094889.
A novel interplay between AR and DAX‐1 controls aromatase expression in estrogen‐dependent cancers
Authors: Lanzino Marilena , Maris Pamela , Sirianni Rosa , Barone Ines , Chimento Adele , Giordano Cinzia , Morelli Catia , Trombino Giovanna Elvi , Bonofiglio Daniela , Catalano Stefania , Andò Sebastiano
Published: June 2021

In situ estrogen production by aromatase is a critical determinant for breast cancer growth and progression while androgens have a protective role in mammary carcinogenesis. Here we demonstrate in breast cancer cells, a novel androgen receptor (AR) mediated mechanism that negatively affects aromatase expression, reducing local estrogen production and cell proliferation. Ligand‐activated AR induces the expression of the orphan nuclear receptor DAX‐1 by binding to a newly identified ARE within the DAX‐1 promoter. In turn, increased DAX‐1 is recruited with the corepressor N‐CoR onto the SF‐1/LRH‐1 site of the aromatase promoter, thereby repressing aromatase expression and activity. Moreover DAX‐1 overexpression induces an apoptotic cell phenotype lying on an enhanced bad/bcl‐2 ratio. This molecular mechanism is reproduced in highly expressing aromatase R2C cells, a model for Leydig tumor, another estrogen‐dependent type of cancer. Interestingly, in vivo studies conducted in testes tissues from Fisher rats, spontaneously developing Leydig cell tumor, confirm the inverse relationship between AR/DAX‐1 and aromatase levels.

In elucidating a novel mechanism by which androgens, through DAX‐1, inhibit aromatase expression in E2‐dependent tumors, these findings sustain the theory of androgen opposing estrogen action, opening new avenues for therapeutic intervention in endocrine‐related cancers.
Natural infection of human adenovirus 36 in rhesus monkeys is associated with gain in body fat, yet, reduction in fasting glucose
Authors: Dhurandhar Nikhil V , Dhurandhar Emily J , Ingram Donald K , de Cabo Rafa , Mattison Julie A
Published: June 2021

Experimental infection of Ad36 increases adiposity, yet improves glycemic control in rodents. In humans, natural Ad36 infection is temporally associated with gain in adiposity and better glycemic control. We determined associations of natural Ad36 infection with changes in adiposity and glycemic control in rhesus monkeys. Monkeys randomized 1y earlier to receive a diet of high fat and sucrose (HFS, N=10) or HFS+resveratrol (RES, N=10), were screened for neutralizing antibodies to Ad36. Ad36‐seropositive (POS; N=5 HFS; 5 RES) and seronegatives (NEG) were compared cross‐sectionally and after 1y follow‐up. In pooled HFS and RES groups, cross‐sectionally, NEG and POS groups did not differ in age, body fat, or fasting glucose (p>;0.9). However, 1y later, fat gain was 510g vs 1245g and reductions in fasting glucose were −3 and −10mg/dL for NEG vs POS, respectively. Gain in body fat and drop in fasting glucose after 1y follow‐up, were significant for POS (p<0.003), but not for NEG. When analyzed separately by diet and seropositivity, only the RES treated POS group showed significant reductions in fasting glucose during 1y follow up (p=0.007). In summary, Ad36 infection is linked with changes in adiposity and glycemic control. Considering effect of resveratrol on reduction in fasting glucose was augmented by Ad36 infection, the interaction may be further evaluated, particularly when studying metabolic interventions.
Dietary Linoleic Acid Elevates Endogenous Endocannabinoids (2‐AG and Anandamide) and Induces Obesity
Authors: Hibbeln Joseph R. , Malde Marian , Osei‐Hyiaman Douglas , Lin Yu Hong , Pawlosky Robert , Madsen Lise , Kristiansen Karsten , Frøyland Livar , Avelheim Anita
Published: June 2021

Suppressing hyperactive endocannabinoid tone is a critical target for reducing obesity. The endocannabinoids 2‐arachidonoylglycerol (2‐AG) and anandamide (AEA) derive from arachidonic acid (AA). Here we posited that excessive dietary intake of linoleic acid (LA), the precursor of AA, would induce endocannabinoid hyperactivity and promote obesity. LA was isolated to reflect dietary increases in LA from 1 percent of energy (en%) to 8 en% occurring in the United States during the 20th century. Mice were fed diets containing 1 en% LA, 8 en% LA, and 8 en% LA + 1 en% eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) in medium‐fat diets (35 en% fat) and high‐fat diets (60 en%) for 14 weeks from weaning.

Results

Increasing LA from 1 en% to 8 en% elevated AA‐phospholipids (PL) in liver and erythrocytes, tripled 2‐AG + 1‐AG and AEA associated with increased food intake, feed efficiency, and adiposity. Selectively reducing LA to 1 en% reversed the obesogenic properties of a 60 en% fat diet. In summary, increasing prevalence rates of obesity 20th century were successfully modeled; elevating dietary LA increased tissue AA and induced hyperactive endocannabinoid status, subsequently resulting in obesity.
Dietary macronutrients modulate the fatty acyl composition of rat liver mitochondrial cardiolipins
Authors: Stavrovskaya Irina G , Bird Susan S. , Marur Vasant R. , Sniatynski Matthew J. , Baranov Sergei V. , Greenberg Heather K. , Porter Caryn L. , Kristal Bruce S.
Published: June 2021

Cardiolipins (CLs) are a family of phospholipids essential for mitochondrial structure and function. The defective remodeling of the CL molecules (i.e., the changing of the fatty acyl [FA] composition) has been linked to human pathology. We examined diet‐mediated CL changes in liver mitochondria from healthy male FBNF1 rats fed 24 different low‐fat, isocaloric diets. Growth rates and mitochondrial respiratory parameters were essentially unaffected, but mass spectrometry‐based, mitochondrial lipidomics profiling revealed up‐regulation of CLs in rats fed saturated or trans fat‐based diets with a high glycemic index. These mitochondria showed elevated monolysocardiolipins and decreased ubiquinone Q9, implying a low‐grade mitochondrial redox abnormality and compensatory CL elevation. Extended analysis of the rat liver mitochondrial CL pools has revealed previously unobserved regulation of CL, suggesting that (i) dietary fats and, to a lesser extent, carbohydrates induce changes in the relative abundance of CL species; (ii) FA incorporation into mature CLs undergoes both positive (~400‐fold) and negative (≥2.4‐fold) regulation; (iii) dietary lipid abundance and incorporation of FAs into both the CL pool and specific mature tetra‐acyl CLs are inversely related, suggesting compensatory regulation.

This study was funded by U01‐ES16048 (BSK, PI) and P30‐DK040561 (W. A. Walker, PI).
Comparison of mass‐action law algorithm‐based pharmacodynamics with the conventional pharmacokinetic studies
Authors: Chou Ting‐Chao
Published: June 2021

Pharmacodynamics (PD) of dose‐effect is governed by the physico‐chemical principle of the mass‐action law. Its systematically derived equations can be deduced to the general median‐effect equation for single entity, and the combination index equation for multiple entity interactions (Chou TC. Pharmacol. Rev. 58: 621–681, 2006, free web access). However, pharmacokinetic (PK) formula are obtained for conventional utilities, such as t1/2, Cmax, Vdis, and clearance rate that are associated with “time”, but not efficacy nor toxicity. PK offers the proper use of drug, e.g., interval/frequency of doses, and precaution for renal or hepatic deficiencies. Contemporary R&D has emphasized PK and ADME, however, rigorous investigation on PD has been grossly neglected which is attributable to the high rate of new drug R&D failure. PD should be the primer. Unfortunately, PD is an underdeveloped area, e.g., bio‐researchers are still drawing the empirical curves to fit numerous data points rather than using a small number of data points to fit the mass‐action law. Furthermore, drug combinations are widely used in treating cancer and AIDS, however, most bio‐researchers are still confused by nearly 20 different definitions for synergy claims without knowing that the mass‐action law algorithm is the key (Chou, Pharmacol. Rev. 58: 621–681, 2006; Scientist 21: 15, 2007; Cancer Res. 70: 440–446, 2010). This confusion leads to inefficiency and waste in research. This author concludes that the mass‐action law PD algorithms and software provide efficient, quantitative econo‐green bio‐research as illustrated in Chou, Integr. Biol. 3: 548–559, 2011 and in Am. J. Cancer Res. 1(7): 925– 954, 2011.