bluuebunny/crossref_metadata_2025 huggingface dataset:
The molecular determinants for ubiquitin NEDD8 discrimination by human USP2
Authors: Chang Shih‐Chung , Chen Jou‐Han , Shin Yung‐Cheng
Published: June 2021

Ubiquitin‐like proteins (Ubls) share very high sequence and structural similarity; but conjugation to a protein leads to distinct biological consequences. Ubls are initially synthesized as precursor forms for processing by Ubl‐specific proteases to expose the C‐terminal di‐glycine motif for conjugation to target proteins. Ubiquitin and NEDD8 share 80% sequence identity, however, each exclusive protease can specifically react with ubiquitin or NEDD8. This work is aimed to study the substrate recognition mechanism of USP (ubiquitin‐specific protease) protein family.

The crystal structure of USP2‐ubiquitin binding complex showed that residues 4, 12, 14 and 72 of ubiquitin were the major interaction sites. It is also noted that these residues are not conserved in NEDD8. By mutating these sites to the corresponding residues in NEDD8, we found that F4K ubiquitin mutant markedly inhibited its hydrolysis by USP2, and T12E and T14E ubiquitin mutants also showed partial catalytic inhibition due to the steric hindrance and electric repulsion between the mutated residue and USP2. Moreover, mutation of residue 72 further inhibited the hydrolysis of T12E/T14E/R72A ubiquitin mutant than T12E/T14E ubiquitin mutant. Because the key amino acids for interacting with residues 4, 12, 14 and 72 of ubiquitin are conserved in the USP protein family, this study suggests a general recognition mechanism by which USP can specifically discriminate between ubiquitin and NEDD8.
Expression of K6W‐ubiquitin in the lens perturbs calcium homeostasis and results in calpain hyperactivation and differentiation abnormality
Authors: Taylor Allen , Liu Ke , Chang Min‐Lee , Shang Fu , Rowan Sheldon , Peng Junmin , Caceres Maria Andrea , Mathias Richard
Published: June 2021

Ubiquitin Proteasome Systems (UPS) control multiple cellular processes but less is known about its roles in regulation of differentiation. The lens must remain clear to function. Precipitation of some lens protein, or their truncated products, result in lens opacification, or cataract. K6W‐Ub mutant enters into conjugates but such conjugates are proteolytically incompetent. There is a dose dependent relationship between expression of K6W‐Ub and the extent of lens aberrations. To initiate elucidation of effects of K6W‐Ub on lens development, we compared protein profiles of lenses from transgenic mice that express wild type ubiquitin (WT‐Ub) or K6W‐Ub. Only ~2% of the proteome was different. Among these are fragmented forms of critical lens proteins including vimentin, γ‐crystallin, filensin, fodrin (spectrin alpha 2), Caprin2 and Tdrd7. In the K6W‐Ub lens, there is a massive accumulation of Ca2+ and hyper activation of calpain. We also observed accumulation of gap junction connexin 43 and diminished connexin 46 levels, consistent with the accumulation of Ca2+ and retention of the epithelial cell type. Corroborating in vitro tests reveal for the first time an association between mutation of K6 on ubiquitin, altered UPS, Ca2+ level regulation, calpain activity, substrate cleavage, and differentiation abnormality. The data are critical to understanding the role of UPS in proper organogenesis.
Serine 32 targeted mutation abolishes Prdx6 trafficking to lamellar bodies in vivo
Authors: Sorokina Elena M. , Dodia Chandra , Yu Kevin , Hong NanKang , Feinstein Sheldon I. , Fisher Aron B.
Published: June 2021

Peroxiredoxin 6 (Prdx6), a protein with both GSH peroxidase and phospholipase A2 (aiPLA2) activities, participates in lung surfactant phospholipid (PL) turnover and protects lung epithelium from oxidative stress. Prdx6 null mice accumulate PL in lung tissue, compatible with the role of PLA2 activity in lung phospholipid turnover. Prdx6 has been localized to cytosolic and acidic compartments (lamellar bodies (LB) and lysosomes) in lung epithelium. Our previous studies indicated that serine at position 32 (S32) is necessary for proper Prdx6 targeting to LB. We have generated a mouse model with an S32T mutation in the Prdx6 LB targeting sequence. Homozygous mice carrying the S32T mutation exhibited phospholipid (PL) accumulation in LB (PL/protein ratio 14.36) vs. WT (PL/protein ratio 9.51). Western blot analysis demonstrated absence of Prdx6 in LBs isolated from S32T mice despite equal amounts of Prdx6 in lung homogenates. Immunofluorescence analysis of mouse lung tissue sections showed co‐localization of Prdx6 with LAMP1, a marker of lysosome‐related organelles, in wild type but not in S32T mouse alveolar type II cells. These findings indicate that S32 is required for Prdx6 targeting to LB in mouse lung epithelium. [HL102016 and HL19737]
Spatial reorganization of yeast moonlighting protein, enolase, in hypoxia to alter carbon metabolism
Authors: Miura Natsuko , Shinohara Masahiro , Tatsukami Yohei , Morisaka Hironobu , Kuroda Kouichi , Ueda Mitsuyoshi
Published: June 2021

Hypoxic conditions have critical effects on physiology of organisms. Using Saccharomyces cerevisiae, we have found that glycolytic enzymes conjugated with GFP formed cellular foci in hypoxic conditions. Enolase is one of foci‐forming glycolytic enzymes under hypoxia and is one of moonlighting proteins. Previously, we have found that N‐terminal part of enolase conjugated with EGFP formed cellular foci [1]. The foci formation of both full‐length and N‐terminal part of enolase was inhibited by single alanine substitution.

In this study we tried to reveal the mechanisms of regulation and the biological function of enolase foci. The formation of enolase foci was inhibited by treatment with N‐acetylcysteine, AntimycinA and by knocking out SNF1. Interestingly, after forming, foci were retained in the cell even in subsequent aerobic culture. Foci‐carrying cells in presence of oxygen could assimilate [U‐13C]‐glucose more rapidly to produce aspartate, compared to cells which have no foci. These results suggest that in response to hypoxia, S. cerevisiae sensed mitochondrial ROS by activating SNF1/AMPK pathway to form complex of glycolytic proteins in cytosol [2]. The formation of foci may facilitate alternative use of carbon sources of hypoxia‐experienced cells.

This work was supported by the Grant‐in‐ Aid for JSPS Fellows [09J02920 to NM].
Vitamin D Supplementation as Influenced by Diabetic Therapies
Authors: Alkharfy Khalid , Al‐Daghri Nasser , Al‐Othman Abdulaziz , Moharram Osama , Alokail Majed , Al‐Saleh Yousef , Sabico Shaun
Published: June 2021

Objective

To the best of our knowledge, no research has determined the effect of vitamin D3 supplementation in conjunction with existing pharmacological and non‐pharmacological approaches to the diabetes mellitus type 2 (DMT2) population. Hence, this study will primarily determine the effect of vitamin D3 supplementation in a cohort of Saudi DMT2 population on different oral hypoglycemic agents and compare them with a non‐DMT2 control.

Methods

A total of 499 randomly selected DMT2 subjects divided into 8 groups [Non‐DMT2 Control = 151; Rosiglitazone (Avandia) = 49; Diet = 15; Insulin alone = 55; Insulin + Orals = 12; Metformin alone = 121; Oral agents combination = 37; Sulphonylurea = 59] were included in this 12‐month interventional study. All subjects were given 2000IU vitamin D3 daily except the control group. Anthropometrics, glucose, lipid profile and 25‐OH Vitamin D were measured at baseline, 6 and 12 months.

Results

Metformin group showed the highest change in vitamin D levels in both 6 months (62.6%) and 12 months (50.6%) as compared to baseline (p< 0.001). No significant changes were observed in the BMI, glucose, total‐ and HDL‐cholesterol in all DMT2 groups. Significant decreases in triglycerides were observed in both the rosiglitazone and insulin + orals group in both 6 and 12 months supplementation (both p‐values < 0.001).

Conclusion

While all groups seemed to increase in levels of vitamin D following supplementation, it appears that DMT2 patients on insulin in combination with other drugs are the group that will benefit the most as compared to other groups in terms of improving cardiovascular risk. Metformin favorably improves vitamin D levels but do not seem to confer other added cardiometabolic benefits.
A Conserved Structural Determinant Located at the Interdomain Region of Mammalian IRE1α
Authors: Xue Zhen , He Yin , Ye Kaixiong , Gu Zhenglong , Mao Yuxin , Qi Ling
Published: June 2021

The UPR sensor inositol‐requiring enzyme 1α (IRE1α) is a bifunctional enzyme containing both kinase and RNase domains that are important for trans‐autophosphorylation and Xbp1 mRNA splicing, respectively, in response to ER stress. However, the amino acid residues important for structural integrity remain largely unknown. Here, through analysis of IRE1α mutants associated with human somatic cancers, we have identified a highly conserved proline residue at position 830 (P830) that is critical for its structural integrity, hence the activation of both kinase and RNase domains. Structural analysis revealed that P830 may form a highly conserved structural linker with adjacent tryptophan and tyrosine residues at positions 833 and 945 (W833 and Y945), thereby bridging the kinase and RNase domains. Indeed, mutation of P830 to leucine (P830L) completely abolished the kinase and RNase activities, significantly decreased protein stability and prevented oligomerization of IRE1α upon ER stress; similar observations were made for mutations of W833 to alanine (W833A) and to a lesser extent for Y945A. Our finding may facilitate the identification of small molecules to specifically compromise IRE1α function.
Stepping into a tense relationship: Mechano‐molecular regulation of cell division by force
Authors: Maresca Thomas
Published: June 2021

To ensure that replicated DNA is equally segregated during cell division, the chromosomes that comprise the genome must interact with and become aligned by the spindle apparatus. An essential protein complex called the kinetochore connects chromosomes to the dynamic microtubule filaments that make up the spindle. Despite the importance of getting it right, cell division is error prone and dividing cells must constantly detect and correct erroneous interactions between kinetochores and spindle microtubules in order to avoid chromosome segregation errors. The goal of my research program is to define and characterize fundamental cell division mechanisms. Towards achieving this goal, we are examining a central yet poorly understood contributor to the process of cell division ‐ force. During cell division forces are generated by microtubule dynamics and molecular motors. Remarkably, the forces produced by motors and microtubules stabilize proper interactions between chromosomes and the spindle through mechanisms that are unclear. We combine high‐resolution microscopy with molecular, cellular and biochemical approaches to characterize how mitotic forces are generated and how the application of force to kinetochores contributes to accurate cell division.
Synthesis and structure‐activity relationship of new cationic lipid derivatives
Authors: Caroyez Benjamin , Lonez Caroline , Ruysschaert Jean‐Marie , Jabin Ivan , Vandenbranden Michel
Published: June 2021

DiC14‐amidine is a cationic lipid that activates Toll‐like Receptor 4 (TLR4), which recognizes lipopolyssacharides. Apart from the presence of hydrocarbon chains, these ligands have so few similarities in terms of size and charge, they are therefore likely to activate the receptor via different mechanisms. Moreover, the role played by the different structural domains of diC14‐amidine remains largely unknown. Therefore, we synthesized a family of derivatives in order to assess the effect of their modifications on TLR4 agonist activity. We observed that a compound containing two non‐natural C13 chains has the highest activity, while an aliphatic chain length of more than 14 carbons has no activation. These results are consistent with the fact that lipopolysaccharides bearing C12 or C14 chains exhibit better TLR4 activity. We showed that a compound for which the secondary amine has been replaced by a methylene group loses all TLR4 activity, suggesting the importance of the amine in the recognition of the ligand. B. Caroyez is recipient of a FRIA fellowship, C. Lonez is a Postdoctoral Researcher (Wiener‐Anspach Foundation) and M. Vandenbranden is a Research Associate at the Belgian Fund for Scientific Research F.R.S./FNRS
Determination of protocatechuic acid from Rubus coreanus fruits and their inhibitory effects of triglyceride accumulation on 3T3‐L1 cells
Authors: Park Hee Sook , Kim Gun‐Hee
Published: June 2021

Obesity is a serious health problem in industrialized countries because it is implicated in various diseases. With the recent increased interest in antiobesity, many investigators are interested in finding phytochemicals such as triterpenoid, essential oil, flavonoid that play a role in this adipocyte differentiation mechanism. Fruits of black raspberry (Rubus coreanus) belonging to the Rosaceae family has been used for centuries as a traditional alternative medicine. This study was investigated determination of protocatechuic acid and their effects of TG accumulation on adipocyte differentiation in 3T3‐L1cells. From this extract fractionated with petroleum ether (A1) and MtBE (A2). We used HPLC to determine the level of protocatechulic acid in A2 fraction from Rubus coreanus. The protocatechulic acid in A2 fraction from Rubus coreanus were confirmed by comparison of the retention times and UV spectra of the samples with those of standard compounds. Thousand ppm of A2 fraction from Rubus coreanus contained 76.64 μM of protocatechulic acid. Anti‐adipogenic effects of A2 fraction from Rubus coreanus were investigated by measuring cell viability and differentiation in 3T3‐L1cells. Treatment of 3T3‐L1 cells with A2 fraction of Rubus coreanus decreased cell viability in time‐dependently. And treatment of A2 fraction inhibited lipid accumulation on adipocytes by 28.8% in 100ug/ml. We also found that containing compounds exerted inhibitory activity in the adipocyte differentiation in this study. Protocatechuic acid which is contained in A2 fraction form Rubus coreanus inhibited lipid accumulation on adipocyte by 26.8 % in 10ug/ml. Thus, Rubus coreanus is a possible candidate for regulating lipid accumulation in obesity.
Novel biodegradable microporous bacterial cellulose scaffolds engineered for bone and cartilage regeneration
Authors: Favi Pelagie , Benson Roberto , Neilsen Nancy , Ehinger Corey , Dhar Madhu
Published: June 2021

The objective of this study was to generate biodegradable, microporous and surface modified BC scaffolds to mimic native bone and cartilage tissues. Microporous BC scaffolds were synthesized and oxidized to render it degradable. To mimic bone tissue, calcium‐deficient hydroxyapatite (CdHAP) was deposited on the BCs. To mimic cartilage tissue, amination and carboxylation of matrices were performed on the scaffolds to simulate the glucosaminoglycans present in the native tissue. Resulting scaffolds were evaluated using EqMSCs. The EqMSCs formed multilayers on the BC scaffold surface. The various forms of BC hydrogel scaffolds exhibited distinct differences on the rate of proliferation and in vitro differentiation. Compared to native BC, non‐degradable modified BCs did not enhance the rate of cell proliferation after 14 days. However, degradable modified BCs did enhance the rate of cell proliferation after 14 days compared to native BC. Although proliferation rate and differentiation potentials varied between the scaffolds, cells were viable and support osteogenic and chondrogenic differentiation on all modified scaffolds. These findings demonstrate that native and specifically modified BC scaffolds are promising constructs for bone and cartilage tissue engineering therapies. This research is sponsored by the National Institute of Health Training Grant (NIH/NIGMS‐IMSD: R25 GM086761).
A Phosphoglycolate Phosphatase from Saccharomyces cerevisiae
Authors: Blake Kimbria Justine , Gehret Austin U. , Puts Regina , Patron Alejandra Rizo , Ramirez Sebastian , Strassner Amanda , Lof Alessandra , Wahler Brandon , Walling Megan , O'Handley Suzanne F.
Published: June 2021

The nitrophenyl phosphatase family of the Haloacid Dehalogenase superfamily consists of a number of phosphatases. The homolog in S. cerevisiae was only known to hydrolyze p‐nitrophenyl phosphate. BLAST searches and alignments indicated that PHO13 from S. cerevisiae was most similar to an algae PGPase. We cloned and overexpressed PHO13 and determined that PHO13 is a PGPase. In photosynthetic organisms, phosphoglycolate is generated in the Calvin cycle; how it is produced in nonphotosynthetic organisms is less clear. Since it a strong inhibitor of triose phosphate isomerase, it must be degraded, regardless of its origin. The PGPase from S. aureus is a virulence factor, indicating its importance. To better understand the role of PGPase in the cell, we are using yeast as a model organism, studying a pho13 knockout mutant and doing complementation studies. Enzymatic characterization, including substrate specificity, pH optimum, divalent metal ion requirements, and kinetics, has been completed, and crystal growth for structural determination is in progress. This research has been supported by an NIH AREA grant, an ASM summer undergraduate research fellowship, and a RIT honors summer research fellowship.
Fast Fix, Fish and Filter, 4Facts strategy to identify real time protein‐protein interactions in situ with few false positives
Authors: Zhu Lisi , Li Menglin , Jia Lulu , Guo Zhengguang , Wei Lilong , Gao Youhe
Published: June 2021

To study dynamic protein‐protein interactions in physiological state, Fast Fix is essential. Formaldehyde cross‐linking can fix the transient and weak protein interactions, but produces great complexity. To reduce the complexity, immunoaffinity purification can Fish complexes of the target protein, but co‐purification by affinity has limited ability in eliminating the nonspecific interactions of beads and antibodies. To Filter the complexes, SDS‐PAGE was used to disrupt non‐covalent bonds thereby eliminated uncross‐linked complexes and at the same time provided molecular weight information for quality control of identified complexes. Only proteins appeared in ranges with molecular weight higher than the sum of their theoretic molecular weights were considered ligands in situ. This strategy tried to combine the advantages of above methods. Proteins which have post‐translational modification can also show up in gels at much higher than that of potential complexes. So the corresponding control was used to eliminate this possibility. In the present study, human blood was added to10% formaldehyde to cross‐link with ligands for five seconds, complexes of albumin and transferrin were purified using their antibodies respectively, then separated in SDS‐PAGE and analyzed by LC‐MS/MS. Proteins identified were further filtered by molecular weight information. Application of this strategy in human blood generated 33 and 24 ligands for albumin and transferrin respectively. Comparing with three previous major albuminome studies, 30% of 33 ligands in our study were also identified in other studies. But only around 15% of their ligands in each of three studies were identified in our study. This 4facts strategy can be used to reveal real time transient and weak protein interactions in situ with less false positives.
Structural basis of human cannabinoid CB1 G protein‐coupled receptor activation
Authors: Scott Caitlin Eileen , Abrol Ravinder , Ahn Kwang H. , Huang Yuehan , Kendall Debra A. , Goddard William A.
Published: June 2021

Transmembrane signal transduction is achieved by activation of G protein‐coupled receptors (GPCRs) like the human cannabinoid CB1 receptor. These receptors exist in an ensemble of conformations, each of which might bind to different signaling molecules. Mutating a single residue, Thr 210, to Ile in the third transmembrane (TM3) domain, makes it more active then WT, whereas mutating it to Ala makes it fully inactive. We used the Gensemble method to predict 3D structures of these receptors. We find conformational differences that explain the CB1 receptor's activation mechanism. These predictions were validated by designing double mutants that were expected to switch the inactive T210A back to WT levels of activation. The 2nd mutation for T210A is predicted to cause an important salt‐bridge between TMs 2 and 6 to break. GTPδS binding assays show a large increase in G protein‐coupling for the double mutants indicating increased activation. We docked known agonists to these receptors and then performed 50 ns of molecular dynamics. The inactive T210A receptor with the docked agonist WIN55212‐2 maintains two stable interhelical salt‐bridges. Similarly, the WT receptor maintains a salt‐bridge between TMs 3 and 6, which suggests that a G protein is necessary to stabilize the active conformation.

This work was funded by gifts from PharmSelex/Accelerator and NIH grants (R01NS071112, R01NS073115, R01AI040567, and DA020763).
Extracts of sweet potato leaves protect β cells from oxidative stress‐mediated injury
Authors: Lin Chia‐Yu , Chiang Tzu‐Yuan , Huang Pei‐Jane , Li Po‐Jun
Published: June 2021

Pancreatic β‐cells are very sensitive to oxidative stress due to relatively low in the expression of antioxidant enzymes, which might play an important role in β‐cell death in diabetes. In the present study, we investigated whether the extracts of sweet potato leaves (SPL) have protective effects against hydrogen peroxide‐induced damage in INS‐1 pancreatic β‐cells. Total phenolic compounds, scavenging activity against DPPH radicals, reducing power method, and ferric chelating ability were evaluated for the antioxidant ability of SPL samples. INS‐1 cells were treated with SPL and/or H2O2 for various concentrations from 2mg/ml to 0.5mg/ml for 24 hours. The reduced cell proliferation after H2O2 toxicity is significantly recovered in the presence of SPL as evaluated by MTT assay (p<0.05). The H2O2‐induced overproduction of intracellular reactive oxygen species is greatly inhibited by SPL extracts (p<0.05), suggesting the free radical scavenging activity of SPL. In parallel, extracts of SPL increase activities of antioxidant enzymes including catalase, superoxide dismutase, and glutathione peroxidase in H2O2 treated INS‐1 cells (p<0.05). In summary, these results suggest that SPL might be used as potential nutraceutical agent which will protect the pancreatic β‐cells from oxidative stress associated with diabetes.
Cytosine DNA Methylation Influences Drug Sensitivity through SugE Expression in Escherichia coli
Authors: Mandarano Alexandra H , Varechtchouk Olga , Simon Robert D , Militello Kevin T
Published: June 2021

In Escherichia coli, the second cytosine in the sequence 5′CC(A/T)GG3′ is methylated by the Dcm enzyme. Our laboratory has previously shown that DNA methylation represses expression of ribosomal proteins. The laboratory is currently investigating other targets of Dcm‐mediated DNA methylation. The sugE gene has been shown to confer resistance to quaternary ammonium compounds, and the sugE gene has one dcm recognition site in its 5′ flanking region, three in the open reading frame and one in the 3′ flanking region. We used qPCR to measure sugE RNA levels at both log and stationary phases in wild‐type and dcm knockout cells. Our data demonstrate that the sugE gene is overexpressed at both phases in dcm knockout cells. SugE expression also increased in the presence of 5‐azacytidine, an inhibitor of cytosine DNA methylation. In order to determine if dcm influences sensitivity to different antibacterial compounds, the sensitivity to ethidium bromide (EtBr) in wild‐type, dcm knockout, and sugE knockout strains was compared using Kirby‐Bauer disc diffusions assays, MIC assays and growth curve analysis. Our data indicate that sugE knockout cells are hypersensitive to EtBr, whereas dcm knockout cells have increased resistance to EtBr. We are currently testing a panel of quaternary ammonium compounds in these assays. In summary, DNA methylation can influence the sensitivity to antibacterial compounds.
Maternal high‐fat diet increases rat hepatic interleukin‐4 (IL‐4) mRNA expression and is associated with decreased DNA methylation in male offspring
Authors: Wang Huan , Zhou Dan , Wang Yijin , Pan Yuan‐Xiang
Published: June 2021

DNA methylation and histone modification are two influential contributors to transcriptional regulation in response to environmental stress, including maternal overnutrition. We observed that a maternal high‐fat (HF) diet in rats caused an inflammatory response in livers of male offspring rats, as measured by the induction of Interleukin‐4 (IL‐4), an anti‐inflammatory cytokine, and this was accompanied by a decreased number of liver inflammatory foci in HF offspring. Methylated DNA immunoprecipitation (MeDIP) results indicated that livers of HF offspring had hypomethylation of specific regions on the IL‐4 gene compared to Control. Methylation‐specific PCR (MSP) confirmed the hypomethylation in the downstream +6 kb region in the HF group, which was significantly correlated with the increased expression of the IL‐4 gene. Moreover, chromatin immunoprecipitation showed an upregulation of acetylation of histone H4 at the same region. The mRNA expression of transcription factor, CEBP/beta was, significantly increased in the HF group compared to Control, and transcriptional factor binding analysis of the selected region within IL‐4 showed strong CEBP/beta binding. In conclusion, maternal HF diet induced hepatic IL‐4 gene expression in male offspring, potentially by decreasing DNA methylation of specific regions as well as through chromatin remodeling.
Regulation of ATP‐dependent Chromatin Remodeling
Authors: Cairns Bradley , Clapier Cedric
Published: June 2021

The objective of our research is to understand how ATP‐dependent chromatin remodeling machines (Remodelers) are regulated. Remodelers contain a catalytic ATPase subunit that conducts DNA translocation, used to pump DNA around the nucleosome. Here, we define and characterize new domains/regions present on the ATPases of the RSC and ISWI remodelers, and show how they regulate ATPase, translocase and nucleosome remodeling activities.

The catalytic ATPase of the RSC remodeler is bound by a heterodimer of actin‐related proteins (ARPs). Here, we present a crystal structure of the ARP module at high resolution. In order to understand how ARPs regulate DNA translocation, we applied genetic, biophysical and biochemical approaches. Here, we reveal a new domain that functions as a negative regulator, restricting ATPase and DNA translocation activities, which is antagonized by the ARP/HSA module. For ISWI remodelers, we reveal an autoinhibition mechanism for ISWI regulation involving an N‐terminal domain, and also define a novel region in the C‐terminus of the ISWI enzyme important for the coupling of energy‐consumption with DNA translocation. Together, these discoveries emphasize the importance of the regulation of ATPase, translocase and remodeling activities by an elaborate network of domains. This work is supported by NIH GM60415 and HHMI.
Latino caregivers are less assertive with family and friends about their child's beverage consumption
Authors: Schilling Elizabeth A. , Quesada Catalina , Ferris Ann M.
Published: June 2021

Caregiver behavioral skills are key to changing children's eating habits. The relationship between demographic characteristics and caregiver assertiveness with family and close friends regarding their child's beverage consumption were examined with baseline data from an intervention study aimed at reducing sugar‐sweetened beverage (SSB) consumption by pre‐school children. Caregivers of preschoolers (n=472) provided demographic information and completed an Information‐Motivation‐Behavioral Skills measure related to child SSB consumption (IMB‐mCSSB). Caregiver assertiveness was defined with 2 items on the IMB‐mCSSB. Caregivers with high assertiveness indicated that “telling [close friends or family] that I do not want my child to have certain drinks” would be “easy” or “very easy.” Logistic regressions showed that although caregiver age, gender, and food security status were not associated with assertiveness, ethnicity was significantly related (p < .0001). Only 57% of Latino caregivers rated the assertiveness items as “easy” or “very easy” compared to 79% of African American, 73% of West Indian, and 76% of White caregivers. These results suggest that culturally sensitive assertiveness training with Latino caregivers may be a helpful component in interventions aimed at modifying child consumption behavior. Funding: USDA/NRI 2008–55215‐19071
Developing Mass in Motion Kids: A multi‐sector intervention to reduce childhood obesity using community‐clinical partnerships
Authors: Blaine Rachel , Taveras Elsie , Davison Kirsten , Gortmaker Steven , Anand Shikha , Falbe Jennifer , Criss Shaniece , Perkins Meghan , Kwass Jo‐Ann , Land Thomas , Smith Lauren
Published: June 2021

As part of a state‐community‐university partnership, Mass In Motion Kids (MiM Kids) was designed as a community‐based intervention to prevent and reduce obesity among primarily low‐income children ages 2–12. Two intervention communities in MA were identified by demographic features and community interest. MiM Kids addresses key behaviors, including screen time, consumption of sugar sweetened beverages, physical activity, sleep duration/sleep quality, and replacement of calorically‐dense foods with fruits and vegetables. Using the Obesity Chronic Care Model as a foundation, literature review and stakeholder feedback informed selection of evidence‐based interventions targeting primary health care (High Five for Kids; Healthy Weight Collaborative), child care (I Am Moving, I am Learning; NAP SACC) and schools/after‐school programs (Food & Fun, Eat Well and Keep Moving, and Planet Health). Environmental policy change (e.g. healthy dining, park space), technical assistance and training, and a youth‐driven communication campaign were also identified as strategies to engage residents. Since MiM Kids was built upon an existing statewide initiative, it is well positioned to be replicated throughout MA if found to be efficacious.

Grant Funding Source: Centers for Disease Control and Prevention: U18DP003370–01
Restaurant meals ‐ almost a full day's worth of calories, fats and sodium
Authors: Scourboutakos Mary J. , L'Abbe Mary R.
Published: June 2021

The objective was to systematically analyze the nutritional profile (calories, fat, saturated fat, trans fat, sodium and cholesterol) of meals from chain sit‐down/family style restaurants. Using the “U of T Restaurant Database”, the nutritional profile of 3507 meals from 19 restaurants was calculated and analyzed. This included all potential meal combinations from restaurants that provided nutrition information online and had 10 or more locations nationally. On average, breakfast, lunch and dinner meals contained: 1129 calories (56% of the average daily 2000 calorie recommendation), 151% of the amount of sodium an adult should consume in a single day (2263 mg), 89% of the recommended daily fat intake level (58 g), 83% of the recommended daily saturated and trans fat intake level (16 g saturated fat and 0.6 g trans fat) and 60% of the daily value for cholesterol (179 mg). More than 80% of meals exceeded the daily recommended intake level for sodium (1500mg). Only 1% of meals had less than 600 mg of sodium, the “healthy level” for meals, according to the FDA. Meals identified by the restaurants as being “healthy” contained on average 474 calories, 13 g of fat (20% DV), 3 g of saturated fat (17% DV) and 752 mg of sodium (50% AI). These data demonstrate that addressing the nutritional profile of restaurant meals should be a major public health priority.

Grant Funding Source: MS: CIHR/PICDP Fellowship, Ontario Graduate Scholarship, ML: U of T McHenry Grant, Canadian Stroke Network
Interrelationships of growth, development and biomarkers of iron status
Authors: Balakrishna N , Radhakrishna KV , Nair K Madhavan , Fernandez‐Rao S , Ravinder P , Hurley K M , Tilton N , Harding K , Reinhart G , Black M M
Published: June 2021

Objective

To study the relationships between a set of observed variables and a set of continuous latent variables among growth, development and iron statues in rural infants using confirmatory factor analysis.

Methods

Z scores of height for age, weight for age and arm mid‐upper arm circumference for age were calculated. Motor, language and visual reception were in 497 infants of 6–12 months using Mullen Scales of Early Learning. Indicators of iron status, hemoglobin, ferritin and sTfR were analyzed. Comparative fit index (CFI), goodness of fit index (GFI), normed fit index (NFI) and root mean square error of approximation (RMSEA) were used for testing the model.

Results

CFI (0.962), GFI (0.966) and NFI (0.935) values met the criteria for an acceptable model fit ¡Ý0.90. The RMSEA obtained was 0.052 which is close to a cut‐off value of ¡Ü0.05 suggesting a good fit. The relationship between development, and indictors of iron status was significant r=0.130 (p<0.05). Fine motor and visual reception showed higher loadings for development. Other contributing variables were found to be weight and sTfR.

Conclusions

The results suggest that the model is valid to establish the interaction between iron status and development. Research support: Mathile Institute & Micronutrient Initiative
Dietary energy density as a risk factor for obesity and cancer
Authors: VERNARELLI JACQUELINE Andrea , Mitchell Diane C , Rolls Barbara J , Hartman Terry J
Published: June 2021

Obesity is an established risk factor for cancer. The objectives of the current study are first to determine whether dietary ED is related to obesity‐related cancer risk factors, and then to evaluate what dietary patterns contribute to this relationship.

Data from a nationally representative sample of 8,550 adults ≥18y who participated in the 2005–2008 NHANES were analyzed. Food energy density (ED, energy per weight of food, kcal/g) was calculated and categorized into one of five categories. Age‐ and sex‐specific quartiles of ED were created to examine the relationship between ED and other obesity‐related risk factors for cancer.

ED was positively associated with abdominal obesity in both men and women in multivariate models. Adults in the highest ED quartile had a 55% increase in risk for abdominal obesity (WC >;88cm in women; 102cm in men) compared to those in the lowest quartile (OR=1.2, 95%CI 1.1–1.4) after adjusting for relevant cofactors. When investigating dietary trends contributing to this relationship, it was observed that lean individuals consume significantly more low‐ED foods (<2.25kcal/g) and fewer high‐ED foods (≥2.25kcal/g) than obese persons.

Overall dietary ED and consumption of high‐ED foods were both positively associated with established obesity‐related risk factors for cancer in a nationally representative sample of US adults.
Harmful Relationship between Alcohol Consumption and Brachial Ankle Pulse Wave Velocity : No Favorable Zone for Vascular Damages
Authors: Kim Mi Kyung , Shin Jinho , Kweon Sun‐Seong , Shin Dong Hoon , Lee Young‐Hoon , Chun Byung‐Yeol , Choi Bo Youl
Published: June 2021

Arterial stiffness and increased intima‐media wall thickness is one of the biggest predictors of cardiovascular disease. We evaluated whether brachial‐ankle pulse wave velocity (baPWV) and common carotid artery intima‐media wall thickness (CCA‐IMT) are correlated with alcohol consumption among men and women aged 40 years and over in a rural area of Korea(2,216 men and 3,428 women). The baPWV was positively correlated with alcohol consumption in men (p for trend <0.0001), but not in women. Age (middle‐aged or elderly) modified the effect of alcohol consumption on PWV and the relation was more evident in the elderly. There was no favorable zone of alcohol consumption in relation to baPWV. CCA‐IMT was likely to decrease with alcohol consumption in both men and women and a linearly decreasing trend for CCA‐IMT with alcohol consumption was found for women, but there was no statistically difference across alcohol consumption groups. In conclusion, there may be not only a linear harmful relationship between alcohol consumption and the arterial stiffness but also neutral effect of alcohol on carotid atherosclerosis without any favorable zone. The effect of alcohol on arterial stiffness may be stronger among elderly men.
Caffeine Intakes from Beverages in the U.S.
Authors: Mitchell Diane C , Hockenberry Jon , Teplansky Robyn , Hartman Terryl J
Published: June 2021

The caffeine intake from beverages was estimated for the US population using the Kantar Worldpanel Beverage Consumption Survey. This survey included a nationally representative sample of 37,815 consumers of caffeinated beverages (≥ 1 yr of age). Respondents completed 7‐day intake diaries including the type, amount, and preparation of each beverage. Parents or guardians completed surveys for children 1–12 yrs of age. Results show that 84% of the US population consumes at least 1 caffeinated beverage per day. The mean daily caffeine intake from all beverages was 165±1 mg for all ages combined. Caffeine intake was highest in the 50–64 yr age group at 223±2 mg and 2.2±0.03 mg/kg body weight. Mean daily intakes were lowest in consumers <6 yrs of age at 36±3 mg and the 6–12 yr age group on a mg/kg body weight basis at 0.9±0.03. The 90th percentile intake was 552 mg for all ages combined, ranging from 182 mg at 6–12 yrs to 651 mg in the 50–64 yr age group. Coffee was the primary contributor to caffeine intakes in all age groups combined but a more significant contributor in adults (>;18 yrs). Carbonated soft drinks and tea beverages were also significant caffeine sources, particularly in the younger age groups. The percentage of energy drink users was low (≤10%) and these beverages were minor contributors to overall caffeine intakes in all age groups. Supported by the International Life Sciences Institute North America; Caffeine Working Group
Development and Validation of Infant Feeding Algorithms Used to Identify Factors Associated with Infant Feeding Practices
Authors: Zhu Yeyi , Hernandez Ladia M. , Dong Yongquan , Mueller Peter , Hirschfeld Steven , Forman Michele R.
Published: June 2021

The objective of this study was to develop and validate infant feeding (IF) algorithms, and identify factors associated with IF practices among 1452 mother‐child pairs from 9 study centers in the National Children's Study Formative Research in Physical Measurements. A test set (N=300) was randomly selected using the remainder as a re‐test set. IF practices including exclusive breastfeeding (XBR), breast‐bottle feeding (BrBot), and exclusive bottle feeding (XBot) were categorized based on maternal recalls for children under six years of age. BrBot was further classified as formula and breast milk (BM) equally, more formula than BM, or more BM than formula. The distribution of XBR, BrBot and XBot did not differ between the test and re‐test sets by age, gender, or ethnicity but differed in one study center. A discriminant function analysis using child and maternal characteristics predicted 89% and 73% of the IF categories in the test and re‐test sets, respectively. Multinomial logistic regression showed mothers born outside the U.S. were more likely to feed by XBR and BrBot than XBot, compared to those U.S. born, adjusted for study centers (N=1452). For BrBot mothers, Non‐Hispanic Whites were more likely to feed more BM than formula than Non‐Hispanic Blacks. The developed and validated algorithms provide unique opportunities to assess associations between these IF practices and health outcomes.

Grant Funding Source: NICHD #HHSN275200800020C
Infant and maternal bone status are associated in lactating adolescents when calcium intake is low but not following calcium plus vitamin D supplementation
Authors: Diogenes Maria Eduarda , Bezerra Flávia F. , Rezende Elaine P , Taveira Marcia Fernanda , Normando Paula , Pinhal Isabel , Donangelo Carmen
Published: June 2021

The aim of the study was to evaluate the influence of calcium plus vitamin D supplementation during adolescent pregnancy on relationships between infant and maternal bone status during lactation. Adolescent mothers (13–19y) with dietary calcium intake ≈600 mg/d, received daily calcium (600 mg) plus vitamin D3 (200 IU) (n=30) or placebo (n=25) from 26 wk gestation until parturition. Infant and maternal DXA scans were done at 5 wk postpartum. Infant measures included total body bone mineral content (BMC), bone area (BA), and bone mineral density (BMD). In mothers, BMD was measured at total body, lumbar spine (L1–L4), total hip and femoral neck. Associations between infant bone measurements and maternal BMD Z scores were evaluated by partial correlation analysis. In the placebo group, infant BMC and BA were negatively associated with maternal total BMD Z score (r = −0.40; P < 0.05 and r = −0.47; P < 0.02, respectively) while infant BMD was positively associated with mothers’ BMD Z scores at all evaluated regions (r ≥ 0.39; P ≤ 0.05). No associations between infant and maternal bone measurements were observed in the supplemented group. Our results suggest that in adolescent pregnancy the calcium demand for neonate bone accumulation is supplied at the expense of maternal bone mass when calcium intake is low.

Financial support: CNPq and FAPERJ (Brazil).
Breastfeeding protection against infant mortality in Mexico, adjusting for reverse causality
Authors: de Cosío Teresita González , Escobar Leticia , González Luz Dinorah , Gatell Hugo López , Hernández Mauricio
Published: June 2021

The effect of breastfeeding (BFing) on the decline of infant (<1yr) mortality (IM) may be overestimated if early neonatal deaths, in infants who cannot breastfeed, are included. Mexican 2012 National Health and Nutrition Survey (NHNS‐2012) data were used. Infant deaths in the last five years were identified by maternal recall (n=1,458 infant mothers, representing, 2,042,011 women). Infants ≥ 1month were considered breastfed (BFed) if mother reported to have BFed her/him for at least 1 month before the survey; infants < 1 month were considered BFed if they received breastmilk for a minimum of 15 days. Mortality was calculated for BFed and not BFed infants in two scenarios: (1) including all infant deaths and (2) excluding 27 (representing 38,289) infants who died and were too sick to breastfeed, to account for reverse causality. Mortality rate ratio (MRR) comparing infants exposed versus unexposed to any BFing, were estimated with Poisson regression. MRR for scenario 1 was 3.01 (CI95=2.73, 3.29) and was reduced to 1.32 (CI95=1.21–1.44) for scenario 2, which accounts for reverse causality. Non‐breastfed infants had 32% higher mortality than breastfed infants, after controlling for reverse causality. Lactation in Mexico is largely unprotected and breastfeeding rates are decreasing, according to NHNS‐2012, though current policy is addressing this decline. These IM data point to the magnitude of the potential benefit neglected due to non‐breastfeeding. Funding for the National Health and Nutrition Survey was obtained from the Mexican Secretariat of Health.
The effect of simulated gut digestion (SGD) of various proteins on cholecystokinin (CCK) and glucagon‐like peptide‐1 (GLP‐1) release in an enteroendocrine cell line (STC‐1)
Authors: Napawan Nida , McGraw Nancy J , Tulk Barry , Krul Elaine S
Published: June 2021

The satiating property of protein is partly due to its ability to stimulate release of gut hormones such as CCK & GLP‐1. STC‐1 enteroendocrine cells were used to determine whether CCK & GLP‐1 release vary by protein source (animal or vegetable) or processing by food enzymes. Proteins were subjected to simulated gut digestion (SGD) using pepsin & pancreatin. Soluble peptide fractions (2 mg/mL) were applied & incubated with STC‐1 cells to evaluate hormone secretion into media. CCK release was increased for the SGD‐proteins (dairy whey protein isolate (WPI), egg albumin, beef protein isolate (BPI), beef plasma protein, & soy protein isolate (SPI)) compared to their intact counterparts (p<0.0004), with SPI (1.6‐fold >; avg.) & BPI (1.4‐fold >; avg.) showing the greatest enhancement (p<0.0001). When BPI, SPI & SPI hydrolyzed by food enzymes (H‐SPI) were compared, the SGD‐proteins showed significantly greater CCK release than before digestion (p<0.001). Only SPI & H‐SPI showed increased GLP‐1 release after SGD (p<0.0001) & SGD‐BPI showed lower GLP‐1 release compared to the other proteins. The average fold stimulation of CCK & GLP‐1, respectively, after SGD was as follows: 1.3‐ & 1.2‐fold (H‐SPI >; SPI); 1.4‐ & 4.4‐fold (SPI >; BPI). In summary, protein‐induced stimulation of CCK & GLP‐1 release is greater for SPI & BPI compared to other sources tested after SGD & is further enhanced for SPI after food enzyme hydrolysis.
Characterization of the ADRP Fatty Acid Binding Site: A Fluorescence Binding and Circular Dichroic Study
Authors: Atshaves Barbara , Lwande Joel , Najt Charles , Gupta Shipra , Moon Kenneth , McIntosh Avery , Kuhn Leslie
Published: June 2021

Although ADRP (adipose differentiation‐related protein, also known as perilipin 2) has been shown to bind fatty acids (FA) and other lipids with high affinity, the structure and location of the FA binding site has not been examined in detail. This is of some importance since ADRP's affinity for lipids and especially FA may be related to lipid droplet formation, triacylglycerol accumulation, and FA uptake and metabolism in the cell. The purpose of the present investigation was to characterize ADRP and several truncated mutants using fluorescence binding assays and circular dichroism (CD) to assess the structural requirements for binding. Results presented here show that removal of 120 amino acids from the N‐terminus, a high homology region common to other lipid droplet proteins increased the FA binding affinity (Kd) 43%. In contrast, deleting 254 residues decreased the Kd 24%. Removal of 173 amino acids from the C‐terminus, however did not significantly change the protein's ability to bind FA. Conformational changes upon binding were confirmed by CD. Taken together, these results indicate that residues 121–254 appear essential for highest affinity binding and show that the ability of ADRP to bind FA may depend on the proper folding of several domains, the partial deletion of which leads to altered, but not ablated ligand binding. This work was supported by NIH Grant DK70965 (BPA).
Treadmill exercise training modulates PCSK9 metabolism in high fat fed mice
Authors: Wen Shin , Jadhav Kavita S , Williamson David L , Rideout Todd C
Published: June 2021

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a novel biomarker of LDL clearance and is a potential therapeutic target of cardiovascular disease risk. Our objective was to examine the effects of aerobic exercise training in modulating PCSK9 metabolism and hepatic sterol regulation in a high fat fed C57Bl/6 mouse. Mice (n=8) were randomly assigned to a high fat diet (HF) or a high fat diet with exercise (HFE) group for 8 weeks. The HFE group was progressively trained 5 days/week on a motorized treadmill (up to 5% grade, 45 min/day, 26 m/min). Endpoint bodyweight was significantly reduced (p<0.05) in the HFE group compared with the HF animals (48.36±1.60 vs 41.13±1.04 g). Aerobic training reduced (p<0.05) plasma LDL/VLDL and PCSK9 concentrations by 41 and 31%, respectively, compared with the HF group. Compared with the HF group, hepatic PCSK9 mRNA was increased (~2 fold, p<0.05) in the HFE group without a corresponding change (p>;0.05) in protein abundance. Aerobic exercise was further associated with an increase in hepatic LDLr mRNA (~1.4 fold, p<0.05) but no change (p>;0.05) in total tissue LDLr protein abundance was observed compared with the HF group. These results suggest that treadmill exercise modulates hepatic and blood PCSK9 metabolism and may underlie exercise‐induced lipid responses in high fat fed C57Bl/6 mice.

Grant Funding Source: UB start up funds