bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nuclear Orphan Receptor COUP‐TFII in Reproduction
Authors: Tsai Sophia Y , Kurihara Isao , Lee Dong‐Kee , Qin Jun , Petit Fabrice , Takamoto Norio , DeMayo Francesco J , Tsai Ming J 
Published: June 2021

COUP‐TFII (Chicken Ovalbumin Upstream Promoter‐Transcription Factor II) is an orphan nuclear receptor that belongs to the nuclear receptor superfamily. Genetic ablation of COUP‐TFII results in early embryonic lethality due to defects in angiogenesis and heart development. COUP‐TFII is expressed in the uterine stroma and is controlled by progesterone‐Ihh‐Ptch signaling. To assess the uterine function of COUP‐TFII, we have ablated COUP‐TFII in the uterine stroma by PR‐Cre. The uterine specific null mutants are infertile and exhibit defects in blastocyst attachment and decidualization. We identified a genetic pathway in which the epithelial Ihh regulates COUP‐TFII to control BMP‐2, which then regulates decidualization. Interestingly, enhanced epithelial estrogen activity, which impedes the maturation of a receptive uterus, was noted in the absence of COUP‐TFII. This finding reveals that COUP‐TFII mediates progesterone signaling to inhibit estrogen activity, and establishes the requirement for epithelia‐stromal interaction during the implantation process. COUP‐TFII is also expressed in the Leydig cells of male mice. Conditional ablation of COUP‐TFII in the male results in infertility due to impairment of spermatogenesis and Leydig cell differentiation. Taken together, COUP‐TFII is a critical player for female and male reproduction.HL 076448, U54 H007495 PJ4, DK 62434
Long–term effects of caloric restriction on total and resting energy expenditure in healthy adults
Authors: Das Sai Krupa , Gilhooly Cheryl H , Golden Julie K , Pittas Anastassios G , Fuss Paul J , Dallal Gerard E , Saltzman Edward , Roberts Susan B 
Published: June 2021

The effects of long‐term caloric restriction (CR) on resting metabolic rate (RMR) and total energy expenditure (TEE) in humans is uncertain. Objective To examine the effects of a 30% CR regimen on TEE and RMR. Methods One year randomized controlled trial of 30% CR in 29 healthy overweight adults (mean±SD, age 35±5y; BMI 27.5±1.5 kg/m2). All food was provided for 6 mo, and self‐administered for the next 6 mo. Outcomes included TEE, RMR, and body fat and fat free mass (FFM). Results Absolute values for RMR decreased during the intervention compared to baseline but measured RMR during CR was not significantly different from RMR predicted from body composition, and RMR changes over time assessed at 3, 6 and 12 mo were not statistically significant after accounting for decreases in body fat and FFM. TEE also decreased significantly with CR but at 12 mo there was a greater reduction in TEE than that anticipated due to the loss of FFM and fat mass (P=0.018). Conclusions This study found no evidence for long‐term metabolic adaptation to weight loss in assessments of RMR, but an adaptive reduction in TEE was observed, which may serve to protect the body against weight loss and contribute to weight regain.Support: NIH grants: NGA‐3U01‐AG20480& #H150001; USDA No. 58‐1950‐4‐401.
Growth and Body Composition of Human Milk‐Fed Premature Infants Provided with Extra Energy and Nutrients Early after Hospital Discharge: One Year Follow‐up
Authors: O'Connor Deborah Louise , Aimone Ashley , Ward Wendy , Vaughan Jennifer , Jefferies Ann , Campbell Douglas M , Asztalos Elizabeth , Feldman Mark , Rovet Joanne , Westall Carol , Whyte Hilary 
Published: June 2021

The purpose of this pilot was to investigate the impact of early post‐hospital discharge supplementation of human milk with a multi‐nutrient fortifier on the growth and body composition of premature infants to one year corrected age (CA). Predominantly human milk‐fed infants (750–1800g birth weight) were randomized to an intervention (n=19) or control (n=20) group at discharge. Infants in the intervention received ~½ of their feedings supplemented with a multi‐nutrient fortifier (24kcal/fl oz, 22g protein/L plus other nutrients) for 12 weeks. Anthropometrics were determined at discharge and at 4, 6 and 12 months CA, and body composition at 4 and 12 months CA using dual energy x‐ray absorptiometry. Intervention infants were heavier, and longer at 12 months CA compared to control infants (p<0.005). Intervention infants born ≤ 1250 g had a larger mean head circumference throughout the first year of life (p=0.0002). Whole body bone mineral content at 4 and 12 months CA was greater in the intervention (p=0.02) but not when controlled for length. In conclusion, adding a multi‐nutrient fortifier to the milk provided to predominantly human milk‐fed premature infants early after discharge results in sustained differences weight, length, and in smaller babies, head circumference for the first year of life.Funded by the Canadian Institute of Health Research(CIHR) and the CIHR Training Grant in Clinical Research
Oxidative Stress in Transgenic Mice with AZT Cardiomyopathy
Authors: Cucoranu Ioan Corneliu , Kohler James , Hosseini Seyed , Johnson David , Green Elgin , Hoying Amy , Russ Rodney , Fields Earl , Wittich Bree , He Stanley , Jones Dean , Lewis William 
Published: June 2021

BACKGROUNDCM and myocarditis occur in HIV/AIDS and symptomatic heart failure is reported in up to 5% of HIV/AIDS patients. Highly active antiretroviral therapy (HAART) significantly reduced morbidity and mortality of AIDS, but has resulted in an increase in cardiac and skeletal myopathies. The thioredoxin (TRX) system is a ubiquitous thiol oxidoreductase system that regulates cellular reduction/oxidation status in the cytoplasm (Trx1) and in mitochondria (Trx2). To identify if oxidative stress is involved in cardiomyopathy induced by the HAART component Zidovudine (AZT) two TGs were employed. One TG exhibits down‐regulated cytosolic thioredoxin in the heart (Tg‐Trx1‐DN) the second TG exhibits overexpression of mitochondrial thioredoxin (Tg‐Trx2).METHODS: TGs and WT littermates were treated with AZT (0.22 mg/day via gavage, n=7). After 35 d, left ventricle (LV) mass and left ventricle end diastolic dimension (LVEDD) were analyzed quantitatively by echocardiography (ECHO; 2D and M mode).RESULTS: In WT, AZT significantly increased LV mass with a mild increase of LVEDD. Both treated and untreated Trx2 overexpressor TGs exhibited LV mass and LVEDD that resembled untreated WT, indicating protection from AZT CM. In contrast, Trx1‐DN mice exhibited baseline left ventricle hypertrophy which was decompensated to cardiac dysfunction with AZT therapy. There was minimal change in LVEDD.CONCLUSIONS: Data suggests that AZT produces cardiomyopathy and oxidative stress is integral to development of AZT cardiomyopathy. Trx2 overexpression protects against AZT cardiomyopathy and Trx1‐DN mice demonstrate increased AZT sensitivity in the heart. Trx2 overexpressors and Trx1‐DN mice together may represent an important model of CM from AZT or oxidative stress.
Regulation of the soluble epoxide hydrolase by peroxynitrite
Authors: Barbosa‐Sicard Eduardo , Froemel Timo , Hammock Bruce D , Fleming Ingrid 
Published: June 2021

Inhibition of the soluble epoxide hydrolase (sEH), which for example converts epoxyeicosatrienoic acids to dihydroxyeicosatrienoic acids (DHETs), has anti‐inflammatory as well as anti‐hypertensive effects. As the enzymatic core of the sEH hydrolase domain contains two tyrosine residues (Tyr383 & 466), we addressed the hypothesis that the activity of the sEH may be affected by oxidative stress, in particular by nitration of Tyr383 & 466.sEH activity was measured in human and murine endothelial cells by LC‐MS/MS and was sensitive to the specific sEH inhibitor 1‐adamantyl‐3‐cyclohexylurea. The importance of the tyrosine residues was demonstrated in that mutation of Tyr383 & 466 to phenylalanine resulted in a complete loss of DHET production. sEH activity in both cell types was significantly (80±6%, P<0.001) attenuated by preincubation with the peroxynitrite (ONOO−) generator 3‐morpholino‐sydnonimine (SIN‐1, 0.5 mmol/L). The latter compound and authentic ONOO− also elicited the tyrosine nitration of the sEH (immunoprecipitation and Western blotting) an effect that was not be triggered by hydrogen peroxide (1 mmol/L). Moreover, in an in vivo murine model, oxidative stress induced by bacterial lipopolysaccharide significantly (20±9%, P<0.01) attenuated lung sEH activity. These data indicate that the activity of the sEH and thus cellular EET levels can be regulated by the tyrosine nitration of the protein.
RNA virus infection affects the location and possibly the function of microsomal epoxide hydrolase
Authors: Duan Hongying , Kobayashi Nobuharu , Takagi Akira , Morisseau Christophe , Hammock Bruce D. , Akatsuka Toshitaka 
Published: June 2021

We have reported that the autoantibody to microsomal epoxide hydrolase (mEH) is induced during the course of hepatitis C and hepatitis A virus infection (J. Autoimmunity 28:7, 2007). The appearance of mEH in sera during the incubation period and the virus specificity of the antibody response suggested that mEH is somehow affected by virus replication rather than by hepatocyte damage. In this study, we have elucidated the effects of RNA virus replication on the location and biochemical function of mEH in cell lines.Human hepatocyte cell lines were infected with various RNA viruses or DNA viruses. They were also treated with poly (I:C) by adding to the medium or transfection. The mEH expression was analyzed by radioimmunoassay, immunofluorescence test, and western blotting. Metabolic activation of a carcinogen by these cells was evaluated by DMBA‐mediated cytotoxicity assay.mEH was found to appear on the cell surface after infection with RNA virus but not with DNA viruses whereas the whole amount of mEH did not change significantly. Such changes could also be induced by poly (I:C) treatment The cells which displayed surface expression of mEH exhibited more than ten‐times higher sensitivity to DMBA‐mediated cytotoxicity.These results suggest that RNA virus infection disrupts the distribution of mEH in the cells and affect the metabolic activation of polycyclic aromatic hydrocarbon carcinogens.
Characterization of the human sEH phosphatase by site directed mutagenesis and LC‐MS/MS analysis
Authors: Cronin Annette , Homburg Shirli , Dürk Heike , Richter Ingrid , Adamska Magdalena , Frere Frederic , Arand Michael 
Published: June 2021

The bifunctional human soluble epoxide hydrolase (sEH) is implicated as a regulator of diverse physiological processes due to the brakedown of arachidonic acid derived signalling molecules like epoxyeicosatrienoic acids (EETs) by its epoxide hydrolase domain. Recently, we discovered that the sEH N‐terminal domain displays a novel phosphatase activity. sEH accepts the generic substrate 4‐NPP as well as lipid phosphates, but the physiological role remains uncertain.The phosphatase domain contains three conserved sequence motifs, including the potential catalytic nucleophile Asp9, and several residues implicated in substrate turnover and/or Mg2+ binding. To enlighten the proposed catalytic mechanism we constructed active site mutants by site directed mutagenesis, which were recombinantly expressed as soluble proteins, purified and analysed for their kinetic properties. An exchange of Asp9, Lys160, Asp184 or Asn189 results in a complete loss of phosphatase activity, emphasising the requirement of these amino acids for catalysis, whereas a substitution of Asp11, Thr123, Asn124 and Asp185 leads to sEH mutant proteins with residual phosphatase activity. To confirm the role of Asp9 as catalytic nucleophile we presently analyze the presumed phosphoester intermediate by mass spectrometry. The dual phosphatase and epoxide hydrolase activities give new insights into the physiological role of human sEH.
Comparison of placenta‐derived MSC (pMSC) with bone marrow MSC (bmMSC) expanded in GMP‐compatible medium
Authors: Pilz Gregor A , Braun Julian , Abele Harald , Schäfer Richard , Aicher Wilhelm K. 
Published: June 2021

Objectives:Human term placenta is a promising source of mesenchymal stromal cells (MSC). After expansion in GMP‐compatible medium containing human‐platelet‐rich plasma, these cells could be utilized in regenerative medicine. In our study we examined the properties of pMSC in comparison to bmMSC.Methods:PMSC were isolated from term placenta and expanded in FCS‐free, GMP‐compatible medium. Characteristic MSC antigen expression of plastic adherent cells was observed by flow cytometry. Differentiation to adipogenic, osteogenic and chondrogenic lineages was detected by cytochemistry and immunostaining. Marker gene expression was measured by quantitative RT‐PCR.Results:PMSC can be expanded efficiently in GMP‐compatible medium while retaining their typical antigen expression and showing differentiation potential comparable to that of BM MSC.Conclusion:Human term placenta can be used as a ethically unobjectionable source of MSC and represent a non‐invasive alternative to bmMSC. The cultivation of MSC in “GMP‐compatible medium” allows rapid expansion to provide the necessary cell numbers for tissue engineering
Alpha‐synucleinopathy is associated with spinocerebellar ataxia type 2 with Parkinsonism
Authors: Takao Masaki , Sakiyama Yoshio , Saito Yuko , Aoyama Masahiro , Shiroyama Harumi , Kurisaki Hiroshi , Ishikawa Kinya , Tsuchiya Kuniaki , Mihara Ban , Murayama Shigeo 
Published: June 2021

Levodopa‐responsive Parkinsonism may be dominant phenotype of patients with SCA2. In general, they have a shorter abnormal expansion of CAG repeats (less than 39) in ATXN2 gene. The aim is to elucidate neuropathologic basis of this variant SCA2 from different Japanese families and give a new perspective on pathogenesis of SCA2 with Parkinsonism.Case 1: He was diagnosed as having Parkinson's disease at age 50. At age 65, he showed cerebellar ataxia and slow eye movement. Case 2: He developed unstable gait at age 40. In addition to the ataxia, he had rigidity, bradykinesia, stooped posture and freezing gait. Genetic analyses of ATXN2 showed an expanded CAG repeat with 22/38 (case 1) and 22/37 (case 2).Neuropathology: There is an atrophy of the pons and cerebellum. The most striking finding was the presence of α‐synuclein immunoreactive Lewy bodies (LBs) and neuritis (LNs) in the locus coeruleus, dorsal motor nucleus of vagus, basal nucleus of Meynert and amygdala. LBs and LNs were also seen in the substantia nigra. The distribution of Lewy pathology was consistent with stage 2 (case 1) and stage 3 (case 2) according to Braak's criteria. The 1C2 antibody depicted intranuclear immunopositive deposits in the various regions. In addition, α‐synuclein deposits were not seen in SCA2 cases without Parkinsonism.We conclude that α‐synucleinopathy is vital pathologic alterations of SCA2 with Parkinsonism. JSPS 19500311
Charge‐driven surface‐colloid interaction for detection biomolecules on microarrays.
Authors: Sun Ye , Golovlev Val 
Published: June 2021

The aim of this study is to demonstrate a new approach for high‐multiplexed analysis of RNA, DNA, and proteins without chemical modification of target molecules for detection. The approach utilizes nanometer‐sized particles, which carry electric charges and are attracted by electrostatic force to target molecules captured on a solid surface. The nano‐particles bind to the surface proportionally to the number of target molecules. Location and surface density of the bound particles can be measured by a low‐cost optical detection system. In our study the charge‐driven particle‐surface interaction was investigated using Gouy‐Chapman‐Stern‐Graham model. The model predicts experimental conditions at which nano‐particles bind selectively to large target molecules and do not bind to small capture molecules present on the same surface. Important implication of this result is the possibility to detect DNA, RNA, and proteins immobilized on a surface with no need for chemical modification or labeling of the target molecules. The detection approach has been demonstrated for gene expression analysis and for protein detection using microarrays.This study is supported by NIH grant 2R44GM074311.
Mutagenesis of Membrane Subunits of the Escherichia coli Complex I
Authors: Michel Jose , Vik Steven B 
Published: June 2021

Complex I plays a major role in the electron transport chains of mitochondria and of many bacteria, but little is known about its mechanism. The bacterial version is considered a minimal version of the mammalian complex I. Oxidation of NADH, reduction of ubiquinone, and proton translocation are three essential functions of Complex I. To identify critical amino acids required for these functions we used site‐directed mutagenesis to create amino acid substitutions in subunits L, M and N. These subunits are the largest subunits in the membrane arm of the complex, and are thought to be involved in proton translocation. Substitutions were found that led to reduced activity, and in some cases to a total loss of activity. In addition, truncations and deletions of individual membrane subunits were constructed and analyzed. In general, function of the complex was lost, but truncation or deletion of subunit L was the least deleterious. In addition, western blots of membrane fractions from the deletion strains indicated that the membrane arm of the complex does not assemble when individual subunits are missing.Supported by a grant from the Welch Foundation.
Apoptosis is the Primary Phenotype Evoked by p190RhoGAP Overexpression
Authors: Ludwig Kirsten , Parsons Sarah J. 
Published: June 2021

p190RhoGAP (p190) is a RhoGTPase activator protein (GAP) that has been shown to regulate actin cytoskeletal dynamics via Rho‐dependent signaling pathways. By enhancing Rho‐mediated hydrolysis of RhoGTP to RhoGDP, p190 negatively regulates association of RhoGTP with its downstream effectors. The p190 gene is located on chromosome 19q13.3, which is mutated in several solid tumor types, suggesting its involvement in malignant transformation. Findings that p190 reduced the incidence of PDGF‐induced gliomas in mouse models further implicate it as a tumor suppressor.To gain insight into its mechanism of action, an overexpression approach was taken. Previous studies in our lab revealed that overexpression of p190 in epithelial cells led to a multi‐nucleated phenotype, suggesting that it negatively regulates mitosis. Chromatin condensation, an apoptotic marker, was also frequently observed. In contrast, overexpression of p190 in fibroblasts led to dendrite formation or chromatin condensation. To further elucidate which cellular processes p190 influenced most, both epithelial cells and fibroblasts were examined for the frequency of the three phenotypes in cells overexpressing p190. The most prominent phenotype in both cell types was apoptosis, as determined by TUNEL staining and caspase 3 cleavage. Real‐time microscopy in epithelials revealed that multi‐nucleation resulted in apoptosis, as did dendrite formation in fibroblasts. In conclusion, we found that overexpression of p190RhoGAP primarily resulted in apoptosis of both cell types, and that multi‐nucleation and dendrite formation appeared to be intermediate phenotypes leading to apoptosis in epithelial and fibroblastic cells, respectively. These results suggest that p190 may utilize different pathways in different cell types to mediate its anti‐tumor effects.National Institutes of Health CA39438 (SJP)
TIM‐1 signaling in T cell activation
Authors: Lin Jean Yean‐Jin , Kane Lawrence P. 
Published: June 2021

T cell immunoglobulin and mucin 1 (TIM‐1) is capable of acting as a co‐stimulatory molecule to enhance T cell activation and to modulate CD4+ T cell responses. Currently, relatively little is known about the signal transduction pathways downstream of TIM‐1 that are responsible for controlling these immune responses. Previous work from this laboratory has demonstrated that src‐kinase dependent tyrosine phosphorylation of the cytoplasmic tail of TIM‐1 is important in transmitting the TIM‐1 response in the T cell. The purpose of this study is to explore additional tyrosine phosphorylated proteins that may be downstream of TIM‐1 signaling. Whole cell lysates from TIM‐1 stimulated D10 cells were probed for tyrosine phosphorylation. This work has suggested a previously unrecognized phosphorylated substrate that can be induced by agonistic anti‐TIM‐1 antibodies. We are currently working to identify this protein as well as to better define the conditions under which it is phosphorylated.Supported by NIAID and NCI.
Effect of Albuterol Isomers on TNF‐alpha; Induced ICAM‐1 and Eotaxin Expression in Human Parasympathetic Neurons
Authors: Nie Zhenying , Jacoby David B , Fryer Allison D 
Published: June 2021

Airway parasympathetic neurons express ICAM‐1 and eotaxin which are required for eosinophil recruitment to the nerves. Major basic protein released by eosinophils blocks inhibitory neuronal M2 muscarinic receptors, increasing acetylcholine release and potentiating reflex bronchoconstriction. TNF‐α induces eotaxin and ICAM‐1 in parasympathetic neurons. Here we test whether albuterol, which is used to treat asthma, changes TNF‐α induced eotaxin and ICAM‐1 expression. Airway parasympathetic neurons were isolated from human trachea and grown in serum‐free medium for 1 week. Cells were incubated with 1uM of either R‐albuterol (the active isomer), S‐albuterol (the inactive isomer) or RS‐albuterol for 90 minutes before addition of 2ng/ml TNF‐α for another 4 hours. In the absence of TNF‐α baseline eotaxin and ICAM‐1 expression was not changed by any isomer of albuterol as measured by real time RT‐PCR. TNF‐α induced ICAM‐1 expression was inhibited by R‐albuterol but not by S or R,S‐albuterol. TNF‐α induced eotaxin expression was not inhibited by any isomer of albuterol. The suppressive effect of R‐albuterol on neural ICAM‐1 expression may decrease eosinophil recruitment to the airway nerves, thereby decreasing reflex bronchoconstriction.Funded by a grant from Sepracor Inc. and grants from NIH: HL55543, HL54659, HL071795, ES014601
Graham Snack Consumption in After‐School Snack Programs Based on Whole Wheat Flour Content
Authors: Sadeghi Leila , Grosser Maggie , Anderson Amy , Reicks Marla , Marquart Len 
Published: June 2021

The 2005 Dietary Guidelines for Americans recommend that children consume at least 3 servings of whole grain foods per day. Data from a pilot after‐school snack program indicated that graham crackers, containing 5g whole wheat per serving, were consumed in the range of 73–88%, similar to the refined grain counterpart. This study examined graham snack consumption containing varying levels of whole wheat flour per serving by school‐aged children. Subjects included ~125 elementary school children (grades K‐6) participating in after‐school snack programs in 4 elementary schools. Over 8 weeks, children were offered graham snacks in a random order containing 5, 8, 12, and 16g of whole wheat flour per 30g serving. Plate waste was used to measure graham snack consumption, as children placed their unfinished snacks in a waste receptacle and subsequent waste was measured after each session. At week 9, a subgroup of children participated in focus groups to provide input about their perceptions for product quality and suggestions for improvement. Consumption for the 5, 8, 12, and 16g graham snacks was 78%, 76%, 85%, and 78%, respectively. This study suggests that these snacks can contribute a significant dietary source of whole grain when offered through an after‐ school snack program.Kraft Foods, Inc.