bluuebunny/crossref_metadata_2025_split huggingface dataset:
Accelerated growth during infancy alters bone characteristics and the activity of mesenchymal stem cells (MSC) in neonatal piglets
Authors: Seabolt Brynn , Li Yihang , Stahl Chad 
Published: June 2021

Rapid growth during infancy has been associated with increased risk of childhood obesity. We examined the effects of increased growth rate caused by overfeeding on growth, bone integrity, and activity of MSC in neonatal piglets. Limit fed (LF) animals received a standard piglet milk replacer at a rate to achieve normal growth, while the overfed (OF) group voluntarily consumed nearly 60% more of the same diet for 21 days. Overfed piglets had higher growth rate (P < 0.001) and feed intake (P < 0.05), and lower feed efficiency (P < 0.001) compared to LF piglets. The OF animals had heavier, larger bones with greater bone mineral content than the LF group (P < 0.05). Surprisingly, MSC isolated from OF animals adopted an adipocytic lineage less readily than those isolated from LF animals after induction with adipocytic media. This was indicated by lower gene expression of PPARG, LPL and AP2 (P < 0.05) as well as by decreased lipid accumulation determined by Oil Red O staining. Increased (P < 0.2) gene expression of Runx2 following osteogenic induction was also seen in MSC from OF pigs. Accelerated growth during infancy led to a decreased response of MSC to an adipocytic environment, as well as an increased response to osteogenic induction. These results indicate the potential for neonatal dietary interventions to influence growth and body composition later in life via programming of MSC.Grant Funding Source: NC Agricultural Research Service
Euglycemia following IED‐like blast exposure
Authors: Scrimgeour Angus Gray , Joosen Marloes J. , Young Andrew J. , van Helden Herman H. 
Published: June 2021

Episodes of hyperglycemia are considered to be a secondary insult in traumatic brain injury (TBI) patients and have been shown to be associated with impaired outcome. Intensive insulin therapy and strict monitoring of blood glucose (BG) during neurointensive care is considered crucial to maintain the brain's glucose demand after brain injury, and to avoid both hypo‐ and hyper‐glycemia. This study investigated glucose levels in peripheral blood in the acute phase following IED‐like blast injury.METHODSTwenty‐two anesthetized adult Wistar rats were exposed to a blast induced by an explosive charge (18g Semtex; 120kPa incident pressure; 12 sec positive phase duration) in a 40×40cm blast‐tube; 22 anesthetized, non‐exposed rats served as Controls. Tail blood was sampled at 24h, 48h and 14 days post‐blast. BG was determined using a commercial photometric analyzer.RESULTSAll blast‐exposed rats exhibited transient bradycardia and bradypnea post‐injury, and loss‐of‐conciousness averaged 3min 20sec. Blast induced deficits in spatial learning and memory, and impaired vestibular motor activity. Post‐blast BG values ranged from 4.8 to 7.3 mmol/l over 14 days. No biochemical hypoglycemia (BG <3.3 mmol/l) was observed. Stress hyperglycemia (BG >;10.0 mmol/l) occurring during an acute illness in those with previously normal glucose tolerance was not observed.CONCLUSIONSBlast‐induced TBI did not induce hyperglycemia in this rat model, and hence there is no support for the use of intensive insulin therapy, in contrast to non‐blast TBI patients with hyperglycemia.Funded by the U.S. Army MRMC and U.S. Air Force MSA.
Analysis of MicroRNA Expression Profile in PBMCs of Hypertensive Patients
Authors: Pinho Maria Joao , Vaz‐da‐Silva Manuel , Soares‐da‐Silva Patricio 
Published: June 2021

MicroRNAs (miRNAs) are endogenous repressors of gene expression and have been reported to play important roles in a number of biological processes. We assessed the miRNA system in Portuguese population of hypertensive subjects in order to investigate the potential of miRNA as biomarkers for essential hypertension (EH). Total RNA was isolated from PBMCs from a total of 66 hypertensive subjects (divided into: group 1, medicated HTA I and II; group 2, medicated HTA III and group 3, nonhypertensive with cardiac disease) and 13 non‐hypertensive subjects. Agilent Human miRNA Microarray 16.0 was used to detect the expression of miRNAs. MiRNA arrays showed that 117, 9 and 21 miRNAs were expressed at different levels in group1, group 2 and group 3, respectively. In addition to several miRNAs previously known to be associated with EH, 4 unidentified miRNA candidates were found to be dysregulated in group 1 samples. Among the novel candidates, the genes hsa‐miR‐4306, hsa‐miR‐146a, hsa‐miR‐146b‐5pn, has‐miR‐22 were upregulated. 14 MiRNAs differentially expressed in group 1 were distributed in 5 miRNA clusters in chromosomes 1, 7, 13 and X. We report four novel miRNA candidates in EH; these miRNA sequences require further validation and functional analysis to evaluate whether they play a role in the pathogenesis of EH.Supported by FCT (PIC/IC/83204/2007)
Regional material heterogeneity in craniofacial cortical bone of the genus Pan
Authors: Gharpure Poorva Hemant , Dechow Paul C 
Published: June 2021

Examination of craniofacial biomechanics necessitates regional material properties data as a foundation. We studied the pattern of material properties in the craniofacial complex of the genus Pan. Our aim was to characterize the elastic properties of the chimpanzee craniofacial cortical bone regionally. Former studies substantiate similarities in patterns of these properties within closely related primate species. Specimens were harvested from the craniofacial cortical bone of five adult chimpanzees. A continuous wave ultrasonic technique was used to measure velocities. Elastic coefficients and technical constants derived from mathematical equations quantified elastic properties. Regional comparisons of data using one‐way ANOVA revealed variations in elastic properties. Significant differences were observed in regional density (P< 0.001). The alveolar region displayed the least thick (mean=1.35 mm) and dense (mean=1630mg/cm3) cortical bone. Results for elastic moduli are significant E1 (P<0.007), E2 (P<0.009) and E3 (P<0.001). Shear moduli were prominent in all of the planes; larger values of G23 indicate stiffer bone. Poisson's ratio was also observed to be significant in v23 (P< 0.019). Greatest anisotropy was seen in the zygomatic arch (E2/E3=0.61). Findings suggest that mechanical properties of cortical bone in the chimpanzee craniofacial skeleton are influenced by its density, architecture and loading patterns in response to functional demands of the cranium.Grant Funding Source: NSF Hominid grant 0725141
The Ubiquitin Conjugating Enzyme UbcH7, controls cell migration
Authors: Whitcomb Elizabeth A. , Le Feuvre Aurelie , Taylor Allen 
Published: June 2021

Post translational modification by ubiquitination can target proteins for degradation, allow the interaction of proteins to form complexes or direct relocalization of proteins to different subcellular compartments. As such, ubiquitin controls a variety of essential cellular processes. Previously we demonstrated that the ubiquitin conjugating enzyme, UbcH7 controls both the entry into and progression through S phase of the cell cycle (Whitcomb et. al 2009 Mol. Biol. Cell v20:1–9). In our search for cell cycle regulatory proteins which might be involved in the UbcH7‐mediated control of the cell cycle, we discovered that UbcH7 affects the level of the cyclin dependent kinase inhibitor, p27, but does not affect the similarly regulated cyclin dependent kinase, p21. Interestingly, depletion of UbcH7 affects p27 levels primarily in the cytosol. While nuclear p27 controls the G1 to S transition of the cell cycle through inhibition of the cyclin dependent kinases, cytosolic p27 plays a role in cell migration. Thus, we asked whether manipulation of UbcH7 also affected cell migration and find that depletion of UbcH7 inhibits cell migration. These results suggest that UbcH7 might be an attractive drug target as it plays roles in controlling both the cell cycle and cell migration.This work was supported by grants NIH EY13250 and USDA 581950–5100‐060–01A to AT.
Conditional protein splicing via disulfide bond formation
Authors: Nicastri Michael C. , Xega Kristina , Reitter Julie N. , Mills Kenneth V. 
Published: June 2021

Inteins are intervening polypeptides that self‐catalyze their own excision from the flanking polypeptides, or exteins, in a process called protein splicing. An intein interrupting the DNA PolII in Methanoculleus marisnigri (Mma) can facilitate protein splicing when over‐expressed in E. coli. However, when overexpressed in gor– E. coli, we isolate mostly unspliced precursor. This precursor can be induced to undergo N‐terminal cleavage upon in vitro incubation with 1,4‐dithiothreitol. Unusual migration on SDS‐PAGE, resolvable by reduction, suggests the presence of a disulfide bond in the unspliced precursor. Mutagenesis analysis indicates that the Cys residues involved in the bond are within the intein, suggesting that this inducible intein may be able to regulate the expression of foreign flanking exteins.This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and by the Camille and Henry Dreyfus Foundation.
Expression and auto‐processing of hedgehog‐like proteins from Brugia malayi and Cryptosporidium
Authors: Cahn Sarah A. , Savidis George , Drago Matthew J. , Reitter Julie N. , Mills Kenneth V. 
Published: June 2021

Hedgehog proteins have an N‐terminal signaling domain and a C‐terminal auto‐processing domain, called the HINT domain (for Hedgehog‐INTein, as it shares sequence and structural similarities to self‐splicing inteins). The HINT domain has a C‐terminal sterolrecognition region (SRR) not shared with inteins. We will analyze the post‐translational cleavage between the signaling and HINT domains from a hedgehog‐like protein from the parasitic nematode Brugia malayi and from two species of Cryptosporidium. We have created E. coli expression vectors that encode fusion proteins of either an N‐terminal His tag or GB1 tag to the full signaling and HINT domains. We are optimizing expression and purification conditions for these proteins, with the aim to study the role of conserved intein residues in the hedgehog‐processing mechanism.This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and the Camille and Henry Dreyfus Foundation.
Splicing of a non‐canonical class three intein from Clostridium thermocellum
Authors: Pusztay Jennifer M. , Drago Matthew J. , Powers Taryn L. , Schufreider Ann K. , Connor Katherine R. , Reitter Julie N. , Mills Kenneth V. 
Published: June 2021

Protein splicing is a post‐translational event by which an intervening polypeptide, called an intein, facilitates its own excision from the flanking polypeptides, called the exteins, and the ligation of the exteins. The first step of protein splicing is an amide to ester or thioester rearrangement of the peptide bond linking the N‐extein and intein, facilitated by the N‐terminal Ser or Cys of the intein. The Clostridium thermocellum TerA protein is interrupted by an intein that lacks the N‐terminal nucleophile. The TerA intein splices as a class three intein by bypassing the first step in the protein splicing mechanism and using an internal Cys to facilitate attack at the N‐terminal splice junction. We have examined the influence of conserved residues on the catalysis of this alternative mechanism.This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and the Camille and Henry Dreyfus Foundation.
Evolution of the respiratory system in Testudines: anatomy and phylogeny
Authors: Schachner Emm R , Lyson Tyler R , Farmer CG 
Published: June 2021

The lung is among the most morphologically diverse vertebrate organs, but the underpinnings (e.g., environmental, phylogenetic etc.) of this diversity are not well understood. We analyzed the pulmonary anatomy of a phylogenetically broad range of morphologically diverse turtles that occupy a wide range of habitats to determine the factor(s) that influence pulmonary anatomy. The anatomy of the primary secondary, and tertiary pulmonary bronchi were visualized as 3D surface models using medical grade computed tomography (CT). Preliminary results indicate that certain aspects of lung anatomy are correlated with carapace morphology and thus ecomorph including: 1) the number of lateral secondary bronchi, 2) presence/absence of ventrally projecting sac‐like secondary bronchi, and 3) the morphology of the tertiary bronchi. This suggests that lung evolution in turtles was influenced by environmental as well as genetic factors. These data also demonstrate that certain aspects of lung anatomy are not reliable phylogenetic characters until the relationship between the respiratory system and the environment in sauropsids is resolved.Grant Funding Source: National Science Foundation, American Association of Anatomists
Role of C‐terminus tail into the ligand binding and releasing mechanisms of Antheraea polyphemus Pheromone binding protein1 (ApolPBP1)
Authors: Mazumder Suman , Katre Uma , Mohanty Smita 
Published: June 2021

In moths, pheromone binding proteins (PBPs) transport the hydrophobic pheromone molecules to the membrane‐bound receptors across the aqueous sensillar lymph. We already showed that recombinant Antheraea polyphemus PBP1 (ApolPBP1) expressed with hydrophobic molecule(s) endogenous to the Escherichia coli which keeps the protein in the ligand‐bound conformation at high pH but switches to the ligand‐free conformation at low pH. For the first time, we showed that the delipidated (without the ligand) ApolPBP1 exists primarily in the free form at all pH levels. We also showed the role of Histidine (His70 and His95) residues presented at one end of the binding pocket. Here we are discussing the role of C‐terminus tail located at the other end of the pocket pocket by solution NMR and binding of 1‐aminoanthracene (AMA) by fluorescence spectroscopy. We found out that C‐terminus truncated proteins showed reduced binding affinity towards the ligand at all pH levels, which indicates that apart from helping in the ligand releasing mechanism, C‐terminus may play a role in ligand binding as well. We also examined the role of three charged residues (Asp132, Glu137 and Glu141) present inside the C‐terminus tail. We discovered that, single mutations (D132N, E137Q and E141Q) and double mutations (D132NE137Q and E137QE141Q) have no effect on the conformation of the protein. They behaved as the wild type protein.This research was financially supported by United States Department of Agriculture PECASE award 2003–35302‐12930, and grant 2011–65503‐20030, National Science Foundation grant IOS‐0628064, and National Institutes of Health grant DK082397 to Smita Mohanty.
Vitamin B12 Supplementation of Bangladeshi Women in Pregnancy and Lactation: Effects on Maternal and Infant Status, Breast Milk and Immunity
Authors: Siddiqua T , Islam S , Ahsan K , Rashid M , Hampel D , Shahab‐Ferdows S , Allen LH , Raqib R 
Published: June 2021

Vitamin B12 deficiency is highly prevalent in Bangladesh and other countries with low animal source food intake. Poor B12 status is associated with several adverse outcomes in pregnancy and infancy, and low breast milk B12; however no B12 supplementation trials have been carried out in pregnant women. In a pilot study, women (n=69, 18–35 y, Hb <115 g/L, 11–14 wk pregnant) were randomized to 250 ug B12/d or placebo through 4 mo postpartum. Both groups also received 400 ug folic acid + 60 mg Fe. Women were immunized with HINI vaccine at 26–28 wk gestation. There were no sig. differences between groups at baseline, when 26% women were B12 deficient (<150 pM), 41% had marginal status (150–221 pM), 26% elevated MMA and 61% tHcy (>;8uM). Supplementation increased plasma B12 and lowered MMA in cord blood and in mothers and infants at 4 mo; it also reduced tHcy in cord blood and infants (all P<0.001) but not in mothers of whom 12% remained elevated (>;12 uM) after supplementation. Hematology, iron status indicators and H1N1 antibody titer did not respond to B12 supplementation. Median B12 in colostrum of the suppl.group increased to 778 vs. 320 pM in the placebo group (P<0.001), and 235 vs. 170 pM at 4 mo (P<0.05) in breast milk. Maternal plasma B12 at baseline remained correlated (r=0.446) with concentrations at 4 mo in spite of supplementation. Maternal and infant B12 status sig. improved by 250 ug/d supplements through pregnancy and lactation.Grant Funding Source: Nestle Foundation, SIDA, and USDA, ARS WHNRC
Why Do Women Stop After Successfully Breastfeeding for Three Months?
Authors: Schwager Haley Nicole , Kennedy Tay , Aubuchon‐Endsley Nicki , Thomas David G. 
Published: June 2021

Although breastfeeding (BF) initiation rates are up, duration of BF remains less than ideal. The objective of this study was to determine when and why a sample of 121 women who had participated in a previous study and were predominately BF when their infants were 3 months old stopped BF. The sample was predominately white, well‐educated and married. The on‐line survey (47% response) asked women to indicate the degree to which 21 factors influenced their decision to stop BF. The survey included open ended questions asking what physicians and communities could do to support BF. Participants were also asked where they received most of their support during their BF experience. Age of infant was the major reason for stopping BF. Two major themes from the qualitative questions were that women wanted access to private spaces to BF their infant outside the home and that husbands were critical supports for BF success. Regional differences in barriers to BF duration need to be understood to meet public health goals. Supported by National Research Initiative Grant 2008–35200‐18779 from USDA National Institute for Food and Agriculture.Grant Funding Source: USDA
Quercetin protects osteoblast development in the presence of oxidative stress in fetal rat calvaria cells
Authors: La Stephanie , Messer Jonathan G. , Hopkins Robin G. , Kipp Deborah E. 
Published: June 2021

Studies examining quercetin (QRC) effects on bone health are limited. The purpose of this study was to examine the extent that QRC impacts osteoblast cell development in the presence of oxidative stress, using osteoblast‐like cells isolated from fetal rat calvaria. At confluence, cells were pre‐treated for 12 hours with QRC aglycone (0 or 20 μM). Cells were then incubated for 4 days with QRC aglycone continued at the same dose as pre‐treatment (0, 20 μM) with or without hydrogen peroxide (H2O2; 0, 300 μM), a known inducer of oxidative stress. There were four groups: untreated Control, QRC alone, H2O2 alone, and QRC/H2O2. Expression of genes related to osteoblast phenotype development (runt‐related transcription factor 2, Runx2; alkaline phosphatase, ALP; type I collagen, COLL; bone sialoprotein, BSP; and osteocalcin, OCN) were analyzed using real‐time quantitative PCR. Additionally, alkaline phosphatase staining was used as an indicator of cells with osteoblastic potential. Results for Controls and QRC treated cells were similar. H2O2 alone suppressed ALP, BSP, and OCN gene expression (p < 0.05) and alkaline phosphatase staining. QRC, in the presence of H2O2, partially prevented the H2O2‐ induced suppression (p < 0.05). These results suggest that QRC is partially protective of osteoblast development in the presence of oxidative stress.Grant Funding Source: USDA Agricultural Research Services
Supplemental ketone metabolic therapy slows tumor growth and increases survival time in mice with metastatic cancer
Authors: Poff Angela M , Ari Csilla , Goldhagen Craig , Seyfried Thomas , D'Agostino Dominic 
Published: June 2021

Metastasis is responsible for 90% of cancer‐related deaths in the U.S. Failure to test therapies in natural pre‐clinical models of metastasis has prevented the effective translation of treatments from lab to clinic. Metastatic cancer cells are highly glycolytic, requiring a large supply of glucose for energy. Mitochondrial dysfunction prevents cancer from efficiently using ketones for energy. Furthermore, ketones inhibit cancer cell proliferation in vitro. The metabolic deficiencies of cancer can be exploited with a ketogenic diet (KD) which lowers glucose and elevates ketones in the blood, a strategy that slows cancer progression in animals and humans. We hypothesized that elevating blood ketones with supplemental ketone administration (SKA) would inhibit cancer progression and enhance efficacy of the KD. We tested this metabolic therapy in the luciferase‐tagged VM‐M3 mouse model of metastatic cancer which spreads naturally in an immunocompetent host, mimicking the natural metastatic phenotype. Mice were fed either ketogenic or standard diet with supplemental ketones (1,3‐butanediol or ketone ester.) Tumor growth was monitored by in vivo bioluminescent imaging. SKA slowed tumor growth and increased survival time by 34–51% when delivered with standard or ketogenic diet. We propose that SKA could enhance the efficacy of standard care and improve the outcome of patients with advanced metastatic disease.Grant Funding Source: Office of Naval Research
The ketogenic diet and hyperbaric oxygen therapy work synergistically to slow tumor growth and increase survival time in mice with systemic metastatic cancer
Authors: Poff Angela M , Ari Csilla , Goldhagen Craig , Seyfried Thomas , D'Agostino Dominic 
Published: June 2021

Cancer energy metabolism is characterized by an abnormal dependence on glycolysis and fermentation for energy which can be exploited therapeutically by decreasing glucose availability to the tumor. The ketogenic diet (KD) is a high fat, low carbohydrate diet which decreases blood glucose and increases blood ketones and has been reported to slow cancer progression in laboratory and clinical settings. Tumor hypoxia is a consequence of abnormal vasculature and promotes cancer progression and further increases glycolytic‐dependency. Hyperbaric oxygen therapy (HBO2T) increases oxygen saturation within the tumor and can offset the cancer‐promoting effects of hypoxia. Since these therapies target overlapping metabolic defects within cancer cells, we tested the combined efficacy of KD+HBO2T on cancer progression and survival in a murine model of metastatic disease. KD alone significantly slowed tumor growth and increased mean survival time by 34%. Interestingly, while HBO2T alone did not influence outcome, combining the KD with HBO2T resulted in a supra‐additive effect, significantly slowing tumor growth and increasing mean survival time by 80% in mice with systemic metastatic cancer. Our results support the potential use of KD and HBO2T metabolic therapy as adjuvant treatments for patients with advanced metastatic disease.Grant Funding Source: Office of Naval Research