bluuebunny/crossref_metadata_2025 huggingface dataset:
Resveratrol enhances the effect of lipopolysacchride on human monocytes THP‐1 pro‐inflammatory cytokine expression
Authors: Feng Li , Huang Haiqiu , Schoene Norberta W. , Yu Liangli , Lei Kai Y. , Wang Thomas T. Y.
Published: June 2021

Resveratrol (Res) is a polyphenolic compound existing in grapes, red wine, and certain fruits that exhibits anti‐tumor and anti‐inflammatory properties. The anti‐inflammatory effect of Res at pharmacological concentrations of >25 μM have been reported, but not at a physiological concentration (~5 μM) normally absorbed through diet. In addition, the mechanisms of action remain largely unknown. We addressed these deficiencies using a cell‐based inflammatory model in vitro. A concentration‐dependent reduction in cell growth was observed in cultured human monocytes THP‐1 cells treated for 48 hours with Res that covered physiological and pharmacological concentrations (≤25 μM). Flow cytometry analysis indicated that 5 and 10 μM of Res induced cell cycle arrest at the S phase. At 25 μM, Res induced both G0/G1 arrest and apoptosis. These effects of Res correlated with up regulation of p53‐independent cyclin‐dependent kinase inhibitor 1A (CDKN1A) mRNA expression. More importantly, Res enhanced the lipopolysacchride (LPS, 10ng/mL)‐induced inflammatory markers IL‐1β, IL‐6, IL‐8, MCP‐1, COX‐2, and TNF‐α mRNA expression in a concentration‐dependent fashion. LPS binds to CD14/TLR4/MD2 receptor complex and promotes the secretion of pro‐inflammatory cytokines. These results suggested that Res exerts a concentration‐dependent effect on immune cell growth and responsiveness toward inflammatory stimuli.

Grant Funding Source : U.S appropriated funds to USDA project number 1235‐51530‐052‐00
Randomized, controlled study demonstrates that About Eating, a web‐based curriculum focused on eating competence increases food resource management skills of SNAP‐Ed eligible women in Pennsylvania
Authors: Patterson Jill , Arnold Kristen , Lohse Barbara
Published: June 2021

About Eating is a 5‐lesson, web‐based program developed to enhance eating competence and related skills of low‐income adults; preview at http://web.survey.psu.edu/AboutEatingPreview. Program impact was assessed in Pennsylvania with 512 (224 control, 288 intervention) SNAP‐Ed eligible, healthy women age 18 to 45 y. Research protocol included: randomization to About Eating or USDA's Click ‘n Go website; online surveys at baseline, lesson completion, and program completion; frequent communications and incentives. Surveys included the USDA Food Security Screener and EFNEP questions. Of study completers (n=302) 60% were SNAP users, 42% had ≤ high school education, 39% were food insecure, and 60% were overweight/obese. Total access time for About Eating was 45 ± 27 min. Reviews were positive; > 99% agreed About Eating was interesting, easy to understand and read. About Eating, unlike Click ‘n Go, increased use of written budgets (P=. 008), “Nutrition Facts” labels (P=.013), meal planning (P=.002), and confidence to manage money for healthful foods (P=.001). Compared to changes in controls, the decrease in likelihood of running out of food at month's end was significantly greater for About Eating (P=.04) and increases in the practice of comparing prices to save money tended to be greater for About Eating (P=.06). About Eating effectively improved food resource management skills of low‐income women. Funded by USDA.
Association between sugar‐sweetened beverage consumption and the metabolically healthy obese phenotype
Authors: Green Angela , Jacques Paul F , Rogers Gail T , Meigs James B , McKeown Nicola M
Published: June 2021

Obesity is typically associated with multiple metabolic abnormalities. However, not all obese individuals exhibit metabolic risk factors. Absence of risk factors among obese individuals has been referred to as the “metabolically healthy obese” (MHO) phenotype. Little information exists regarding possible dietary differences between obese individuals with and without these metabolic abnormalities. The purpose of this study was to determine if sugar sweetened‐beverage (SSB) consumption was associated with the MHO phenotype among obese, non‐diabetic adult members of the Framingham Offspring Cohort. We classified MHO as BMI≥30 with none of the 5 following metabolic criteria: insulin resistance, hypertension, elevated fasting glucose and triglycerides, and low HDL cholesterol. Overall prevalence of MHO was 10.1%. Prevalence of MHO was 2‐fold greater among individuals in the lowest quartile of SSB consumption compared to those in the highest quartile (15.5% vs 7.8%, P=0.02) after multivariate adjustment. Our findings suggest that lower SSB consumption is associated with healthier metabolic profiles in obese individuals.

Grant Funding Source : USDA ARS (contract number 58‐ 1950‐7‐707.Number), NHLBI (contract number NO1‐HC‐25195Number)
Women's employment impacts household food expenditure patterns over time in rural Bangladesh
Authors: Mehra Sucheta , Klemm Rolf D. W. , Labrique Alain Bernard , Rashid Mahbubur , Shamim Abu Ahmed , Christian Parul , West Keith Parker
Published: June 2021

In resource‐poor settings, income earning women are hypothesized to have greater control of household expenditures and may prioritize the purchase of more or better quality food. This 4 year longitudinal analysis evaluated the impact of women's formal, part‐time employment with a research project in rural Bangladesh on monthly per capita food expenditure. Women were categorized as unemployed (n=182), first time employed with the project (n=279), first time employed elsewhere (n=184) and transitioning in employment to the project (n=291). Change in women's contribution to household income differed (0%, 37%, 4% and 5% respectively; p<0.001). Monthly per capita total food expenditures increased across time (Tk 382.6 to Tk 1921.3, p<0.001) and differed by group (p<0.05), while the food share of income, similar across groups at baseline (p=0.15), decreased considerably and unequally across groups (p<0.001) 4 years later. Though absolute per capita expenditure on food categories was not different, the share of food expenditure on rice, meat, fish, eggs, dairy, vegetables, fruits and snacks varied by group at baseline, shifted over time and differed in the rate of change by group, which will be presented. Rice expenditure shares increased uniformly but meat, eggs, dairy and fruit were prioritized differently by different categories, indicating that women's income influences household food choices.

Funded by USAID and the Gates Foundation.
No effect of pH on in vitro digestion of carotenoids from sweet potatoes and mandarin oranges
Authors: Burri Betty J. , Devin Coline M. , Rajaonary Maelle , Roman Maxine , La Porte Delphine
Published: June 2021

Beta‐carotene (BC) and beta‐cryptoxanthin (CX) can form vitamin A physiologically. Recent observational studies suggest that CX has greater bioavailability than BC from most diets. This means that CX‐rich foods such as mandarin oranges might be better than expected sources of vitamin A. We hypothesized that CX was more easily digested from its major food sources, possibly because these foods are acidic. We used a static in vitro digestion model to compare the apparent bioavailability of CX from mandarin oranges to BC from orange‐fleshed sweet potatoes (OFSP) at several physiological pHs. The model included saliva, gastric fluid, duodenal fluid, and bile components. BC and CX concentrations were measured by reversed‐phase HPLC or by spectrophotometry at 450 nm. BC from both varieties of boiled OFSP had very low bioavailability (0.65 – 1.08%), comparable to those reported for OFSP without fat in the literature. CX from mandarin oranges had approximately 3X better bioavailability (2.21 – 4.25%). Changes in saliva, gastric, and bile fluid pH did not change bioavailability significantly for either mandarin oranges or OFSP. Our results support the hypothesis that CX is more bioavailable from a major food source (mandarin oranges) than BC is from orange‐fleshed sweet potatoes (OFSP), a common intervention food used to increase VA status. Research was supported by USDA ARS in‐house funds.
Behavioral intervention among early adolescent girls improves bone mass after 18 months; however lactose maldigestion is still a barrier for calcium intake
Authors: Lee Yujin , Boushey Carol J , McCabe Linda D , McCabe George P , Novotny Rachel , Van Loan Marta , Read Marsha , Going Scott , Weaver Connie M , Savaiano Dennis A , investigators ACT
Published: June 2021

A targeted behavioral intervention to improve calcium intake and bone mass was conducted at middle schools in 5 states (473 girls of Asian, Hispanic, or non‐Hispanic White heritages, 10 –13 y). Lactose maldigestion (LM) was determined by breath hydrogen. Perceived milk intolerance (PMI) and calcium intake were assessed by questionnaire and bone mineral content (BMC) was determined by dual energy x‐ray absorptiometry. Multiple linear regression was used to examine the relationship among intervention, PMI, LM, calcium intake and BMC while adjusting for covariates. During the intervention period, there was an attenuated reduction in dietary calcium intake in the intervention group, compared to the control group (−74 mg, −177 mg, respectively). From 0 to 18 mos, a significant increase in total hip BMC was observed in the intervention group. At the end of the intervention (12 mo), dietary calcium consumption had increased among lactose digesters (LDs) and decreased among LMs. Additionally, significant increases in spinal BMC and total hip BMC were observed among LDs compared to LMs (p=0.01, p=0.04, respectively). A targeted behavioral intervention in middle‐school aged girls influenced dietary calcium intakes and bone mass. LM is a barrier to calcium intake in this population and may need alternative interventions.

Grant Funding Source : USDA
The role of cellular junctions in neural crest cell emigration
Authors: Taneyhill Lisa Anne
Published: June 2021

Neural crest cells are a transient population of migratory cells that differentiate to form a wide array of derivatives in vertebrates, including the craniofacial skeleton, peripheral nervous system and skin pigment cells. As such, defects in neural crest cell formation can have drastic developmental consequences and lead to various human congenital and hereditary malformations, diseases and cancers. Premigratory neural crest cells initially reside in the dorsal region of the embryonic neural tube and become motile by undergoing an epithelial‐to‐mesenchymal transition, a process characterized by cytoskeletal rearrangements and the loss of intercellular contacts in order to facilitate their emigration out of the neuroepithelium. Using the chick embryo as a model system, we have performed depletion and overexpression experiments to examine how perturbation of cellular junction components affects neural crest cell development. Our studies reveal that components of cellular junctions in premigratory neural crest cells must be precisely modulated to permit both appropriate neural crest cell migration and subsequent differentiation into developing cranial ganglia. These results have direct implications for craniofacial development and should provide additional insight into the etiology of various neural crest‐derived diseases, disorders and cancers. This research is supported by grants from the NIH (R00HD055034) and NSF (IOS‐0948525).
Tanshinones inhibit androgen‐dependent prostate cancer via downregulation of aurora A and suppression of androgen‐receptor signaling
Authors: Zhou Jin-Rong , Gong Yi , Abdolmaleky Hamid , Blackburn George
Published: June 2021

The objective of this study was to evaluate the effect of tanshinones on androgen‐dependent prostate cancer. Tanshinone I (T1), tanshinone IIA and cryptotanshinone (CT) significantly inhibited proliferation of androgen‐sensitive LNCaP cells in part via induction of apoptosis and cell cycle arrest at G0/G1 phase associated with and down‐regulation of survivin and aurora kinase expression and interference of androgen signaling pathway. Knockdown of aurora A by siRNA significantly reduced the growth‐inhibitory activity of tanshinones, suggesting aurora A as a functional target for tanshinones. Without changing the androgen receptor level, tanshinones dramatically down‐regulated gene expression of prostatic specific antigen and Nkx3.1, but upregulated maspin gene expression. The animal study using an orthotopic prostate tumor mouse model further confirmed that CT and T1 treatment significantly reduced tumor growth associated with induced prostate cancer cell apoptosis, reduced prostate cancer cell proliferation and tumor angiogenesis, down‐regulated prostate cancer cell survivin and aurora A protein expression, without altering food intake or body weight. Our results suggest that CT and T1 may be efficacious and safe candidate agents for the prevention and therapy of androgen‐sensitive prostate cancer. Supported by Idea Award PC073988 (Department of Defense).
Poor positioning, decreased prolactin levels, and low milk output associated with early cessation of exclusive breastfeeding in obese women
Authors: Hauff Laura E , Demerath Ellen W.
Published: June 2021

High prepregnant maternal body mass index (BMI) is associated with reduced breastfeeding duration. The objective of this research was to examine how maternal obesity might impact early cessation of exclusive breastfeeding (EBF) by exploring key aspects of early breastfeeding success: prolactin values, milk output, and mother‐infant positioning during the first week postpartum. In this prospective study, data were collected from 51 primiparous women from Minnesota. Multivariate models were used to determine relationships between maternal obesity and early cessation of EBF via a variety of breastfeeding outcome variables. Among overweight and obese women, 35% stopped breastfeeding exclusively by 2 weeks postpartum compared to 4% of normal BMI women (P < 0.05). Greater milk output at 1 week postpartum reduced the risk of early cessation of EBF (OR 0.41, P < 0.05). Milk output was significantly associated with the predictor variables of basal prolactin concentration within 48 hours postpartum (P < 0.05) and Mother‐Baby Assessment (MBA) score (a measure of efficient positioning) at 1 week postpartum (P < 0.0001). Prolactin levels (P < 0.05) and MBA scores (P < 0.05) were in turn negatively associated with maternal obesity. These findings suggest that low milk output, influenced by decreased prolactin levels and especially poor positioning, may play a role in early cessation of EBF among obese women. Funded by NSF.
Relationships between emotional eating and coronary heart disease risk factors in college students
Authors: Arts Jennifer , Fernandes Jill , Schembre Susan , Hirshberg Shira , Lofgren Ingrid
Published: June 2021

Emotional eating (EE) is hypothesized to have a negative impact on diet quality and body weight that could affect chronic disease risk. Our aim was to determine if EE was associated with diet quality, physical activity (PA), and coronary heart disease (CHD) risk factors among 261 freshmen at a Northeast university. EE was scored with the Weight Related Eating Questionnaire. Three, 24‐hour food recalls were collected to compute Dietary Approaches to Stop Hypertension (DASH) diet scores. Moderate‐vigorous PA (MVPA) was assessed by the International PA Questionnaire. CHD risk factors were measured after a 12‐hour fast. Most participants had a normal body mass index (BMI) (73%; 23.3 ± 3.8 kg/m2), were female (67%), and ranged in age from 18–21 y. Males and females differed significantly (p < 0.01) on all variables except for glucose and low‐density lipoprotein cholesterol concentrations. By multiple regressions, which included gender, we observed borderline significant associations between EE and MVPA (β = −0.12, p = 0.07), and with BMI (β = 0.12, p = 0.06) and diastolic blood pressure (β = 0.12, p = 0.07). Waist circumference was significantly associated with EE (β = 0.16, p < 0.01). However, DASH diet scores and MVPA failed to explain the observed associations between CHD risk factors and EE. Findings confirm associations among EE and some CHD risk factors that appeared independent of diet quality and PA.

Research funded by the USDA

Grant Funding Source: USDA
Vitamin D regulates apoptosis in adipocytes via Ca2+ signaling
Authors: Sergeev Igor N
Published: June 2021

Modulation of apoptosis is emerging as a promising strategy for prevention and treatment of obesity because removal of adipocytes through this process will result in reducing body fat and long‐lasting maintenance of weight loss. Effects of vitamin D on apoptotic cell death are mediated via multiple signaling pathways that involve common regulators and effectors converging on cellular Ca2+ (Sergeev, 2009). The vitamin D‐regulated Ca2+‐dependent apoptotic molecular targets in adipose tissue have not been identified. We investigated mechanism of 1,25(OH)2‐vitamin D3 (1,25D)‐induced apoptosis using mouse 3T3‐L1 adipocytes. The results obtained demonstrated that 1,25D induces, in concentration‐ and time‐dependent fashion, the apoptotic Ca2+ signal ‐ a sustained, prolonged increase in concentration of intracellular Ca2+ ‐ in mature, accumulating lipids adipocytes. The 1,25D‐induced increase in cellular Ca2+ was associated with activation of the Ca2+‐dependent μ‐calpain and the Ca2+/calpain‐dependent caspase‐12. The activation of these proteases was sufficient for effecting morphological and biochemical changes attributed to apoptosis. These findings imply that the 1,25D‐induced cellular Ca2+ signal can act as an apoptotic initiator that directly recruits Ca2+‐dependent apoptotic effectors capable of executing apoptosis in adipose tissue. Targeting of the adipocyte Ca2+ signaling with vitamin D may represent an effective, safe and affordable approach to the prevention and treatment of obesity.

Supported by USDA 2009‐35200‐05008 and USDA SD00H325.
Association between Vitamin D Metabolites in Fat Tissue and Serum 25‐hydroxyvitamin D in Overweight and Obese Adults
Authors: Piccolo Brian D , Gertz Erik , Thomas Anthony P , Keim Nancy L , Adams Sean H , Seyoum Elias , Dolnikowski Gregory , Van Loan Marta D
Published: June 2021

Cholecalciferol has been measured in human white adipose tissue (WAT), but little is known about the relationship between the other circulating vitamin D metabolites and WAT. We measured concentrations of 25(OH)D and 1,25(OH)2D in subcutaneous fat tissue from 20 overweight and obese subjects participating in a weight loss intervention (Van Loan, 2011). WAT biopsies were obtained pre‐ and post‐intervention from the gluteal region and were analyzed for 25(OH)D and 1,25(OH)2D by LC/MS/MS (Blum, 2008). Serum 25(OH)D and 1,25(OH)2D were measured by RIA. Mean adipose 25(OH)D concentrations were 2.24 ± 1.0 ng/g pre‐ and 2.39 ± 1.1 ng/g post‐intervention, but 1,25(OH)2D concentrations were undetectable. Pre‐ and post‐intervention WAT 25(OH)D concentrations were correlated (r = 0.5482, P = 0.0185) and there was a significant positive association between subcutaneous fat 25(OH)D concentrations and serum 25(OH)D concentrations (r = 0.5140, P = 0.0011) across a range of body weight and adiposity. This suggests that similar to the parent compound, 25(OH)D deposition in human WAT is directly dependent on circulating 25(OH)D. The active compound, 1,25(OH)2D, may not be stored in appreciable amounts. Funding: National Dairy Council; USDA, ARS, WHNRC; Dairy Council of California; CTSC, University of California (1M01RR19975‐01), National Center for Medical Research (UL1 RR024146)

Grant Funding Source : Dairy Research Institute
Relationship between brain lutein (L) and zeaxanthin (Z) and retinal L and Z in humans
Authors: Vishwanathan Rohini , Wittwer Jonas , Schalch Wolfgang , Johnson Elizabeth J
Published: June 2021

Background

L and Z preferentially accumulate in the neural retina, as macular pigment (MP), and also in the brain tissue. Given the predominance of L in adult and pediatric brain tissues and the significant association of L with cognitive health in older adults, it is imperative to have a tool to assess brain L and Z status.

Purpose

To evaluate the correlation between brain L and Z and retinal L and Z in humans.

Methods

Voluntarily donated brain tissue and matched retina were obtained from a brain bank which adheres to strict consent and confidentiality procedures. Decedents were men and women aged >50 y who either had normal cognitive function or Alzheimer's disease. Tissues were analyzed using standard lipid extractions and reverse phase HPLC. SPSS version 19 was used for statistical analyses.

Results

Total L and Z in the occipital cortex of the brain was positively related (r=0.865, n=3) to total L and Z in the retina in a bivariate analysis. Due to small sample size the ‘p’ value was non‐significant. Tissues (occipital cortex, hippocampus and retina) from seven more decedents are currently being analyzed.

Conclusions

As MP can be measured non‐invasively in humans, it could be used as a biomarker to assess brain L and Z status.

Support: USDA# 1950‐51000‐065‐08S, DSM Nutritionals

Grant Funding Source : USDA# 1950‐51000‐065‐08S, DSM Nutritionals
Cadavers, death anxiety, and the early development of the medical double‐consciousness
Authors: Martiny Amy , Witmer Lawrence M
Published: June 2021

The objective of this research is to examine the effects of cadaver use on medical student socialization. Medical school exposes students to cadavers, bringing them face to face with death and dying and requiring them to perform acts (dissection) that in any other context would be taboo or even criminal. Student responses to these stressors can be heavily influenced by the curriculum, lab setting, and faculty, impacting the development of their “medical double‐consciousness” (partitioning the humanity of the individual from the scientific needs of the patient). An immersive anatomy course provided a uniquely controlled setting for this study in that it is short, unopposed by other courses, and is the students’ first academic learning activity. It thus provides advantages over previous studies that looked at anatomy courses of long duration and interspersed with other activities that present confounding variables. Data were gathered largely using participant observation and interviews. The preliminary results of this research project identify the variables that impact a student's personal and professional life, such as death‐anxiety coping mechanisms, acquired social roles at the dissection table (cutters, readers, floaters, etc.), and general attitudes prior to, during, and after their gross anatomy lab experiences. Funding from NSF IOS‐1050154, NSF IOS‐0517257, and OU‐HCOM.

Grant Funding Source: National Science Foundation
Lysine rich C‐terminal domain of Ku protein from Mycobacterium smegmatis regulates its mode of DNA interaction
Authors: Kushwaha Ambuj Kumar , Grove Anne
Published: June 2021

Ku protein binds DNA double strand breaks and is an integral component of the non‐homologous end joining (NHEJ) pathway of double strand break DNA repair. Sequence alignment of mycobacterial Ku proteins have shown that Ku proteins from most of the soil dwelling mycobacterial species like M. smegmatis have low complexity sequence at their C‐terminus, which is characterized by lysine‐rich PAKKA repeats. Using electrophoretic mobility shift assays, we have shown that both full length M. smegmatis Ku and truncated Ku (TKu), lacking the C‐terminal PAKKA repeats bind DNA non‐sequence specifically irrespective of the kind of ends (blunt or cohesive) and overhangs (3′ or 5′). Also, by competition assay we have shown that full length M. smegmatis Ku and TKu binds supercoiled DNA and linear DNA with equal preference. Further, we observed that removal of the lysine rich extension from M. smegmatis Ku increases its binding affinity and the binding site size. Altogether, our findings suggest that the C‐terminal PAKKA repeats of Ku protein plays a regulatory role in its interaction with DNA. Research supported by National Science Foundation.
Structure function differentiation of truncated hemolysin A using multi‐angle light scattering
Authors: McDermott James Joyce , Weaver Todd Michael
Published: June 2021

In this study, the role of molecular organization was investigated as related to hemolysin A (HpmA265) template‐assisted behavior. Previous studies have shown that full‐length HpmA can be activated through interaction with purified HpmA265 in template‐assisted fashion. HpmA265 was analyzed via template‐assisted hemolysis, circular‐dichroism, size‐exclusion chromatography and multi‐angle light scattering (SEC‐MALS) before and after tryptic digestion. The results from these investigations were utilized to determine the absolute molecular weight of the template‐assisted active species. The pre‐tryptic digestion data reported a heterogeneous mixture of HpmA265 monomer and dimer. Quantitative hemolytic assays have determined the monomer to be the molecularly active species. Subsequent tryptic digestion, followed by SEC‐MALS, generated a homogenous dimeric species with enhanced template‐assisted behavior and thermal stability. From these data, a new model for template‐assisted activity has been developed where exposed, homogenous, and monomeric on‐edge beta‐strands of HpmA265 facilitate the activation of full‐length HpmA. These findings are significant and may apply to other beta‐helix protein dependent disease states like Alzheimer's, Parkinson's and transmittable prion disorders. National Science Foundation Grant (MCB0744754) and (MCB1050435) supported the work.
Stability Contributions of Hydrogen Bonds in the Third Circuit of Hemolysin A from Proteus mirabilis
Authors: Mauseth Matthew A , Weaver Todd Michael
Published: June 2021

In this study, the role of unique intra‐molecular hydrogen bonds at the third circuit of truncated Hemolysin A (HpmA265) was investigated. HpmA265 harbors a beta‐helix structure, hallmarked by wrapping of parallel, beta‐strands around a central and largely hydrophobic core. The tight packing of beta‐strands in the beta‐helix pattern provides a stable structure via inter‐strand hydrogen bonding networks. A series of site‐directed mutants within the three beta‐arcs of the third circuit have been designed and engineered to investigate the structural and functional contributions of hydrogen bonding in HpmA265. Glycine 148 at the third circuit allows the formation of a carboxy‐amide bridge connecting beta circuits two and four. Replacement of Gly148 with a lysine residue has revealed important thermal stability differences as determined by circular dichroism (CD) spectroscopy data. National Science Foundation awards MCB0744754 and MCB1050435 supported the research.
The Role of Methionine Residues in Activity and Subunit Communication in Malate Dehydrogenase
Authors: Billue Farren , Bell Ellis
Published: June 2021

Malate Dehydrogenase is a dimeric enzyme that catalyzes the oxidation of malate to oxaloacetate in a reaction involving both a hydride transfer [to NAD] and a proton abstraction, and involves subunit interactions in activity and regulation [by citrate]. Little is known concerning the mechanism of communication between active sites in the dimer. The subunit interface contains three conserved methionine residues [M62, M64 & M91] in close proximity to residues from the opposing subunit [P59, L63, Y273 & I88 as well as between M66 on one subunit and M62 & M91 on the opposing subunit]. These residues together with a Methionine close to the active site [M128] have been mutated using QuikChange mutagenesis to a variety of different amino acids to test their role in subunit communication and catalytic activity. Proteins are expressed, purified using Ni‐NTA affinity chromatography and characterized by MALDI‐tof. Circular Dichroism in conjunction with thermal melts has been used to examine both stability and folding cooperativity using a 4 parameter Hill equation to analyze the ellipticity at 222nm as a function of temperature. Initial rate kinetic studies demonstrate changes in both Vmax and Km of various mutants as well as indicating changes in citrate regulation of the protein.

This work is supported by NSF Grant MCB 0448905 to EB and an HHMI summer stipend to FB
Collaborative Center for Chemical and Biological Mass Spectrometry
Authors: Sauer Jon , Lawrence Jim
Published: June 2021

Chemical and biological investigators increasingly depend on data from mass spectrometers to answer complex questions in their research. However, high resolution mass spectrometers are difficult to obtain and operate. Alternatively, access to high resolution mass spectrometers can be obtained through academic core facilities, but this involves other obstacles. We have an Agilent Technologies 6520 QTOF LC MS/MS mass spectrometer capable of producing high resolution MS/MS data and providing this data to outside research groups. We use these collaborative opportunities to develop and improve our methods for sample analysis as well as train undergraduate students in sample handling, MS/MS operation and data analysis methods. Currently we are capable of identifying small organic molecules as well as identification of proteins from SDS‐PAGE gels. We are in the initial phases of experimentation with ITRAQ isobaric peptide labeling procedures coupled with 2‐D liquid chromatography. By forming collaborative projects with other research groups we provide our collaborators with MS/MS data that they would be unlikely to otherwise obtain, we educate the next generation of MS/MS scientists and we have the opportunity to further develop methods for unique samples in a collaborative environment. This work was supported by the National Institutes of Health and the National Science Foundation.
Orf6: Biochemical Characterization of a Putative Thioesterase from a Polyunsaturated Fatty Acid Synthase
Authors: Rodriguez-Guilbe Maria M , González-Méndez Ricardo , Wymore Troy , Screiter Eric , Baerga-Ortiz Abel
Published: June 2021

Polyunsaturated fatty acids are made in deep‐sea organisms by the activity of a polyketide synthase multienzyme. The final step in the biosynthesis of polyketides and fatty acids is catalyzed by the activity of the thioesterase (TE) domain, which cleaves the final product off of the carrier protein. However, no TE domain has been identified in any of the known PUFA synthase clusters. We propose that orf6 gene is a candidate TE, since it is conserved among other bacteria with PUFA synthase clusters and is adjacent to the PUFA synthase cluster in Photobacterium profundum. We have cloned, expressed and purified Orf6 protein with the aim of characterizing the protein structurally and functionally. The Orf6 structure, determined by X‐ray crystallography at 1.0Å resolution, demonstrates a hot‐dog fold similar to the 4‐hydroxybenzoyl‐CoA TE family. Results from enzyme assays show that Orf6 has no activity toward long‐chain fatty acids, including the expected full length product eicosapentaenoic acid attached to a CoA carrier. Additional efforts include in vitro assays using CoA thioesters of short‐chain carboxylic acids as well as aromatic carboxylic acids. Structural and mechanistic knowledge of this enzyme will be important for the development of the PUFA synthases as a platform for the production of fatty acids. This work was funded by grant CHE0953254 from the NSF and MBRS‐RISE Program (R25GM061838) of the UPR‐MSC.
Dietary protein is inversely associated with sarcopenia, sarcopenic‐obesity, and impaired gait speed in US adults: NHANES 1999–2002
Authors: Zanovec Michael , OˈNeil Carol E , Fulgoni Victor L , Nicklas Theresa A
Published: June 2021

As the American population begins to age, nutritional interventions that mitigate the associated losses in lean body mass and physical function will become increasingly important. The NHANES 1999–2002 database was utilized to determine if protein intake was associated with the risk of sarcopenia, sarcopenic‐obesity, and impaired gait speed in adults 50+ y (n=4,010) with reliable 24‐hr dietary recall data, dual‐energy X‐ray absorptiometry measures, and that completed a timed 20‐ft walk test. Covariate‐adjusted logistic regression was used to determine odds of excess body fat percentage (% BF; gender‐specific 75th percentile), sarcopenia (gender‐specific 25th percentile of appendicular lean mass index [ALMI] in kg/m2), sarcopenic‐obesity, and impaired gait speed (<0.8 m/s) by quartiles of total daily protein consumption. Mean values for % BF, ALMI, and gait speed were: 36.5±0.2%; 7.3±0.0 kg/m2, and 1.03±0.01 m/s. Mean daily protein intake by quartiles were: 35.3±0.4 g (0.49±0.01 g/kg); 58.8±0.2 g (0.80±0.01 g/kg); 79.1±0.2 g (1.03±0.01 g/kg); and 121.8±1.2 g (1.47±0.02 g/kg), respectively. Compared to the lowest quartile, subjects in the highest quartile had significantly reduced odds of excess body fat (OR=0.67; 95% CI:0.48–0.93), sarcopenia (OR=0.65; 95% CI:0.43–0.99), sarcopenic‐obesity (OR=0.28; 95% CI:0.13–0.63), and impaired gait speed (OR=0.49; 95% CI:0.33–0.73). Results suggest a beneficial role of dietary protein in reduced risk of sarcopenia, excess % BF, and impaired gait speed in older adults.

Grant Funding Source: Dairy Research Institute and USDA
Quantitation of alpha‐tocopherol metabolism in humans
Authors: Novotny Janet A. , Chuang Jennifer C. , Fadel James G. , Holstege Dirk M. , Furr Harold C. , Clifford Andrew J.
Published: June 2021

To improve the understanding of vitamin E kinetics, a study was conducted in which 12 healthy adults (6 men, 6 women) ingested 1.81 nmol of [5‐14C]‐RRR‐α‐tocopherol and provided serial blood samples for at least 70 days and serial urine and fecal samples for at least 20 days after the dose. Carbon‐14 enrichment of plasma, red blood cells, urine, and feces was determined by accelerator mass spectrometry. Data were analyzed by compartmental modeling. Results revealed that α‐tocopherol exists in plasma in at least 4 distinct kinetic compartments, including chylomicrons, lipoproteins, CEHC, and a very quickly absorbed form. Results also revealed that α‐tocopherol rapidly binds to red blood cell membranes in vivo. The model indicated the presence of several tissue pools with different turnover rates, including a slow turnover tissue pool which may represent deposition of vitamin E in adipose tissue. Mean absorption (±SD) was 81%±6%, and absorption ranged from 73% to 93%. Mean RRR‐α‐tocopherol loss (±SD) via urine was 2.2 μmol/d (±2.0), and loss via feces was 7.5 μmol/d (±3.7). The half‐life of RRR‐α‐tocopherol in the deep tissue pool was on average 203 days. This work was funded by the USDA and NIH.
Energy and nutrient contribution and diet quality of breakfast patterns in children 2–18: National Health and Nutrition Examination Survey 2001–2008
Authors: OˈNeil Carol E. , Nicklas Theresa A. , Fulgoni Victor L.
Published: June 2021

Breakfast meals have not been well characterized. NHANES 2001–2008 data were used to identify breakfast patterns in children 2–18 years (n=14,200). Cluster analysis generated 12 distinct breakfast patterns—including no breakfast. Energy, added sugars, saturated fat, sodium, fiber, vitamin D, calcium, and potassium intake, and diet quality (determined using the Healthy Eating Index [HEI]) were determined for each pattern. For the patterns, energy and intake of added sugars, saturated fat, and sodium consumed at breakfast ranged from 0 (no breakfast) to 584 kcals (meat/grain/juice), 6 tsp (soft drink/juice/grain/potatoes), 10 g (meat/grain/juice), and 1178 mg (meat/grain/juice), respectively. Intake of fiber, vitamin D, calcium, and potassium (nutrients of public health concern) ranged from 0 (no breakfast) to 4.5 g (cooked cereal/milk/juice), 4.3 mcg (ready‐to‐eat cereal [RTEC]/whole milk), 395 mg (RTEC/reduced fat milk), and 627 mg (meat/grain/juice), respectively. HEI for breakfast alone ranged from 0 (no breakfast) to 50 (cooked cereal/milk/juice), and for the day from 45 (meat/grain/juice) to 56 (cooked cereal/milk/juice). Different breakfast patterns had better nutrient intake and diet quality than others. Education is needed to improve breakfast patterns in terms of nutrients to limit and those of public health concern. Supported by USDA and Kellogg's Corporate Citizenship Fund.
Bioavailability and pharmacokinetics of whole grain phytochemicals in overweight/obese adults
Authors: McKay Diane L. , Chen C-Y. Oliver , Gómez-Juaristi Miren , Blumberg Jeffrey B.
Published: June 2021

Whole grains contain micronutrients and phytochemicals as well as fiber. While research suggests that fiber is a major contributor to the health benefits associated with whole grain consumption, other ingredients concentrated in the outer bran layer, e.g., alkylresorcinols, lignans, phenolic acids, phytosterols, and tocols, may also play an important role. To determine the acute bioavailability and pharmacokinetics of the major phytochemicals found in barley and oats, we conducted a randomized, 3‐way crossover trial in 13 healthy subjects, age 40–70 y with BMI 27–35.9. After a 2‐d run‐in period following a diet low in phytochemicals, subjects were randomized to receive muffins made with either 48 g whole oat flour, whole barley flour, or refined wheat flour + fiber (control), with a 1 wk washout period between each intervention. Blood and urine samples were collected at specified time points before and for 24 h following the consumption of each test food. Preliminary results (n=6) indicate the rank order for maximal plasma concentration (Cmax) of ferulic acid, the predominant phenolic acid, was oats>barley>control and time to reach Cmax (Tmax) was 3.6–4.3 h. Minimal changes in plasma phytosterols and tocols were observed, likely due to their low content in these flours. These data suggest that barley and oat phytochemicals are bioavailable. (Supported by USDA and Kellogg Co.).
Clofibrate increases in vivo fatty acid oxidation by neonatal pigs
Authors: Bai Xiumei , Xi Lin , Drayton Josephine , Ji Cheng , Odle Jack
Published: June 2021

To test the effect of clofibrate on the fatty acid (FA) oxidation in vivo, newborn pigs received 5 mL of vehicle (2% tween 80) with or without clofibrate (75 mg/kg bw) once daily via intragastric gavage for 3 days. Total FA oxidative capacity was measured in respiration chambers after gastric infusion of piglets (n=5/treatment) with isoenergetic amounts of [1‐14C] labeled triglycerides (TG) ‐‐ either tri‐C18:1 (3.02 mmol/kg bw0.75) or tri‐ C22:1 (2.46 mmol/kg bw0.75). Total expired 14CO2 was collected and quantified at 20‐min intervals over 24 h. Hepatic in vitro FA oxidation was determined simultaneously using [1‐14C] labeled C18:1 and C22:1. The average 24 h accumulative [1‐14C] TG oxidative utilization (% of energy intake/kg bw0.75) tended to increase with clofibrate supplementation (P<0.14), but there was no difference in the peak utilization rate. The maximal extent of tri‐C18:1 oxidation was 61 % greater than for tri‐C22:1. Hepatic FA oxidation in vitro increased significantly with clofibrate. The relative abundance of mRNA increased 2–3 fold for hepatic PPARα and its target genes (fatty acyl‐CoA oxidase (FAO), carnitine palmitoyltranserase (CPT) I and CPT II) in the pigs given clofibrate. The increase in mRNA enrichment by clofibrate was greater for CPT I than for ACO. In conclusion, clofibrate may improve in vivo FA oxidative utilization in neonatal pigs. Supported by CSREES, USDA NRI program award 2007‐35206‐17897.
Early and more variable growth velocities improved linear growth in Peruvian infants
Authors: Iannotti Lora , Zavaleta Nelly , Leon Zulema , Huasaquiche Clara , Caulfield Laura
Published: June 2021

This study aimed to investigate: growth velocity patterns in Peruvian infants; determinants of velocities during critical periods; and anthropometric outcomes at 12 mo predicted by velocities. From 1995–1997, an RCT of maternal zinc supplementation was conducted in a periurban slum area of Lima. 579 infants were followed monthly through 12 mo on a range of anthropometric measures. We examined 3 types of velocity variables: incremental velocity (1 mo, 2 mo, and 6 mo); 2) proportional changes (% of total size gained/mo); and 3) individual velocity variability (SD of child incremental velocities). Mean within‐child SD of weight velocity was 0.42 kg (±0.12). Growth velocities in month 1 and variable weight velocity positively predicted attained height and weight by 12 mo in multivariate regression analyses. Panel regression by generalized least‐squared of height and weight velocities confirmed the exponentially decelerating pace of growth through infancy and the importance of birth size in driving this trajectory. Arm fat area was a significant determinant of length gain in the same month. Boys grew more rapidly than girls in the first half of infancy, but not in proportion to the total incremental gain. This study contributes evidence to support the importance of early growth velocities and greater degrees of weight gain plasticity for attained height and weight. The study was funded by USAID and NIH.
Silencing the estrogen receptor promoter using DIF‐1, a naturally occurring differentiation molecule of the cellular slime mold Dictyostelium discoideum
Authors: Bratton Melyssa , Stutts Whitney , Pandey Arvind , Burow Matthew , McLachlan John
Published: June 2021

Drugs and environmental chemicals exhibit estrogenic activities by various signaling pathways. Recently, we have shown that pesticides like DDT activate cell signals involved in estrogen receptor (ERα) coregulator recruitment. We have also shown that plant compounds extracted from soy are potent antiestrogens. In the present study we employ a naturally occurring compound secreted by the slime mold Dictyostelium discoideum to probe the basis of estrogenicity. Differentiation inducing factor 1 (DIF‐1) is a morphogen and mitogen in the slime mold. Our objective was to determine the estrogenic activity or lack thereof of DIF‐1 in a model system (ERα (+) breast cancer cell line MCF‐7). Using colony assays, we showed DIF‐1 blocks MCF‐7 cell growth. ERE‐luciferase assays showed DIF‐1 inhibited ERα activity. Quantitative PCR revealed DIF‐1 inhibited the transcription of the ERα regulated genes stromal derived factor 1 (SDF‐1) and progesterone receptor (PR), and Western blot analysis showed DIF‐1 downregulated total ER. Finally, an ER promoter assay was used to show DIF‐1 partially blocks transcription of ERα. This represents a novel mechanism for anti‐estrogenic action in mammalian cells. Our results present DIF‐1 as a novel ER antagonist which will shed light on the mechanism of action of estrogenic compounds. This work was supported by the Office of Naval Research, grant number N00014‐06‐1136.
The role of muscarinic receptors in prefrontal excitability and fear extinction
Authors: Santini Edwin , Sepulveda-Orengo Marian T. , Porter James T.
Published: June 2021

Fear extinction enhances the intrinsic excitability of infralimbic (IL) prefrontal neurons. We recently showed that IL excitability is modulated by M‐type K+ channels. Given that muscarinic receptors (MusRs) inhibit M‐channels, cholinergic inputs to IL may play a role in controlling IL excitability and fear extinction. To test this hypothesis, we combined patch‐clamp recordings and fear conditioning. In brain slices, muscarine increased the number of neuronal spikes by reducing the M‐current indicating that MusRs enhance the intrinsic excitability of IL neurons. Next, we showed that systemic injection and intra‐IL infusion of the MusRs antagonist, scopolamine, prior to extinction training impaired recall of extinction 24‐hrs later. Finally, systemic injections of the muscarinic agonist, cevimeline, facilitated recall of extinction. Together these findings suggest that cholinergic inputs to IL play an important role in consolidation of fear extinction and that muscarinic agonists might be useful to facilitate extinction memory in patients with anxiety disorders. Supported by NSF grant IOS 0842159 to JTP, RCMI Behavioral Core Facility 5G12RR003050‐26 and MBRS RISE 1R25GM082406 to MS.
Cross sectional anatomy of the femoral diaphysis in mice bred for high levels of voluntary wheel running
Authors: Coats Brittney Racheal , Middleton K. M. , Kelley S. A. , Garland T.
Published: June 2021

Artificial selection for increased levels of voluntary activity in mice has resulted in differences in whole bone morphology. To study evolutionary adaptation and phenotypic plasticity of bone in response to increased levels of activity, we compared cross‐sectional geometry of the femoral mid‐shaft in 59 male and female mice from three lines: randomly bred controls, high‐runner mice (HR), and HR mice expressing the Mini‐Muscle (MM) phenotype. Half of each group was either allowed or denied wheel access for ten weeks. From histological sections, we measured several relevant biomechanical traits. Male and female mice generally followed similar trends, although female mice differed significantly in cross‐sectional area among lines while males did not. HR mice in both sexes had the thinnest cortical bone, the largest medullary diameters, and, consequently, the greatest resistance to torsional loads. MM male and female mice exhibited more elliptical femoral mid‐shafts (with males differing significantly), with greater resistances to torsion. We conclude that bone micro‐structure responds both evolutionarily and plastically to increased loading. Future studies will evaluate rates of bone growth resulting in the observed differences. Supported by NSF IOB‐0543429 to TG and NIH 1F32AR053008‐01 to KMM.
The biomechanical effect of the bipartite zygoma: A Finite Element Analysis of the impact of an auxiliary facial suture in the midface
Authors: Unger Evelyn , Wang Qian
Published: June 2021

Typically the zygoma is a single facial bone bounded by sutures where it abuts the maxilla, frontal, sphenoid, and temporal bones. Rarely, the zygoma is transected by an extra suture arising perpendicularly from the facial aspect of the maxillozygomatic suture and running dorsolaterally to the zygomatic aspect of the temporal fossa giving the bone a bipartite morphology (Incidence of 2.5–6.5% in human populations). In the midface, which is normally under heavy stresses during feeding, the biomechanical effect of bipartite zygoma has so far not been explored. We hypothesized that the bipartite zygoma condition would alter the pattern of stress distribution in the face during mastication. The bipartite zygoma condition was created in a Finite Element Model of a skull from a Tufted Capuchin monkey (USNM518342), and the model was analysed and compared to a model with typical zygoma condition. Results revealed that the bipartite morphology disrupts the normal inferosuperior stress flow from maxilla to frontal through the zygoma. The superior half of the zygoma in the upper face was spared high stress flow at the expense of the inferior half of the zygoma in the midface. In this way, the stresses incurred during normal activities (i.e. feeding) were shunted from the upper facial regions to the relatively gracile yet already heavily stressed temporal arm of the zygomatic arch, suggesting that the bipartite morphology is less optimal than the typical case. How the temporal arm of the zygomatic arch adapts in this situation needs further studies.

Grant support: NSF HOMINID BCS‐0725183 and NSF BCS‐0100825 (CT images of USNM518342 courtesy of Dr. James Rossie. www.digimorph.org).