bluuebunny/crossref_metadata_2025_split huggingface dataset:
Changes in Cellular Nucleic Acid Binding Protein with Aging and Degenerative Disease
Authors: Webb Robin , Weidner Adam M. , Holler Christopher J. , Beckett Tina L. , Murphy M. Paul 
Published: June 2021

Background: The ZNF9 gene on chromosome 3 encodes the Cellular Nucleic Acid Binding Protein (CNBP), a ubiquitously expressed protein that is highly conserved among vertebrates. An expansion in the first intron of CNBP results in Myotonic Dystrophy Type II, and it is a potential modifier of β‐secretase (BACE), the rate‐limiting event in the formation of the Aβ peptide, implicated in Alzheimer's disease.Objective: To determine if there are changes in CNBP with aging or degenerative disease, and begin elucidating the effect on BACE.Methods: AAV2 was used to overexpress CNBP in cell culture and in mouse skeletal muscle. Both were evaluated by ELISA, western blot, and RT‐PCR. CNBP levels were evaluated across the murine lifespan.ResultsThere is a significant increase in CNBP murine muscle and brain with aging. Three‐fold overexpression of CNBP was achieved in vivo, leading to an increase in BACE1 protein (p<0.01) and increase in Aβ (p<0.01). In culture, increased BACE1 was detectable after 24 hours. The steady state amount of BACE1 mRNA did not change in vivo or in vitro after CNBP AAV2 treatment.Conclusions: These data support the hypothesis that CNBP regulates BACE1, and that this effect is largely post‐transcriptional or translational. Future studies will elucidate the effect of CNBP knockdown on BACE1 and Aβ related pathology. Supported by NIH/NINDS R01 NS058382 and NIH/HL T32 086341.
Role of brain nitric oxide in cardiovascular control of frogs
Authors: Zena Lucas A. , Gargaglioni Luciane H. , Bícego Kênia C. 
Published: June 2021

The aim of the present study was to determine if nitric oxide (NO) acting on the brain of frogs presents an inhibitory effect on arterial blood pressure (ABP) and heart rate (HR) by reducing the sympathetic activity dependent on α and β adrenergic receptors. Thus, body temperature and ABP was measured by a telemetry device implanted into the abdominal cavity of american bullfrogs (250–350 g BW), with the catheter of the device inserted into the left aortic arch for ABP measurements. A guide cannula was implanted into brain lateral ventricle for injections of L‐NMMA (non selective NO synthase inhibitor) or mCSF (mock cerebrospinal fluid). A catheter was inserted into the femoral vein for injection of adrenergic antagonists (prazosin: α1; sotalol: β) and agonists (phenylephrine: α1; isoproterenol: β) or Ringer solution. Animals (n= 6) were maintained at 25oC during experiments. L‐NMMA increased (P< 0.05) ABP by 7.5±1.3 mmHg compared to mCSF (−0.5±0.7 mmHg) and did not change HR (delta HR: −1.37±1.8 bpm and −0.9±0.6 bpm, L‐NMMA and mCSF respectively). The pre‐treatment with prazosin, but not with sotalol, attenuated the hypertensive effect of L‐NMMA. The effectiveness of the α and β adrenergic antagonists was confirmed by the blockade of the respective adrenergic agonists. We conclude that NO seems to act on the brain of frogs as a hypotensive agent via a mechanism dependent on α, but not β, adrenergic receptors in the periphery.Supported by FAPESP and CNPq.
Protective role of PPARγ receptors against vascular dysfunction associated with insulin resistance
Authors: El‐Bassossy Hany , Fahmy Ahmed , Abo‐Warda Shaimaa 
Published: June 2021

Here we investigated the protective effect of PPARγ receptor stimulation from vascular complications associated with insulin resistance (IR).IR was induced by fructose (10% in drinking water) and left for 12 weeks. PPARγ was stimulated by either rosiglitazone or pioglitazone (5 or 10 mg/kg respectively) in the last 6 weeks. Then, non invasive blood pressure (BP) was recorded. Rings of isolated thoracic aorta were used to measure the responses to phenylephrine (PE), KCl, acetylcholine (ACh) and sodium nitroprusside (SNP) relaxants and to study NO generation and intracellular calcium.IR was associated with elevated systolic and diastolic BP while, both rosiglitazone and pioglitazone significantly reduced the elevation in BP. IR increased response of aorta to PE and KCl, decreased response to ACh without affecting response to SNP, while both rosiglitazone and pioglitazone prevented the impairment in response to KCl and PE but not to ACh. IR was accompanied with reduced ACh stimulated NO generation but exaggerated intracellular calcium increase in response to KCl while both rosiglitazone and pioglitazone restore normal KCl stimulated increase in intracellular calcium but did not affect NO generation.In conclusion, PPARγ stimulation protects from IR induced elevation in BP through preventing impairment in vasoconstriction and calcium movement.
Induction of non‐cardiogenic pulmonary edema following cerebral hypoxia in a canine model
Authors: Khademi Shadi , Frye Melinda , Jeckel Kimberly , Irwin Dave , Hamilton Karyn , Monnet Eric 
Published: June 2021

Neurogenic pulmonary edema (NPE) is a non‐cardiogenic pulmonary edema associated with stroke, traumatic brain injury and seizure with high morbidity and mortality, yet the underlying mechanism(s) is unknown.HypothesisWe hypothesize that cerebral hypoxemia induces pulmonary edema in the presence of systemic normoxia.MethodsUsing an updated Moss's model for induction of cerebral hypoxia, 8 Walker hounds (25–30kg) were randomly assigned into cerebral hypoxemia (SaO2 ~55%) with systemic normoxia (SaO2 ~90%) (CH; n=4), cerebral and systemic hypoxemia (SaO2 ~ 50%) (GH; n=2), and cerebral and systemic normoxia (SaO2 ~90%) (Con; n=2). In this model, femoral venous (hypoxic) or arterial (normoxic) perfusate was delivered from a cardiopulmonary bypass circuit to the brain. Outcomes included the wet to dry lung weight ratio as an index of pulmonary edema, hemodynamic measurements, intracranial pressure (ICP) and cerebral blood flow.ResultsWet to dry weight ratio was increased in the CH and GH groups compared to control. We observed no changes in hemodynamic parameters in CH dogs, with the exception of an increase in ICP. Dogs with GH exhibited increased cardiac output, pulmonary artery pressure and cerebral blood flow, with decreased systemic blood pressures; no increase in ICP was observed.ConclusionBrain hypoxemia, in the absence of systemic hypoxemia, induces pulmonary edema in this canine model. DARPA N66001‐10‐C‐2134.
Perinatal hypoxia increases KCNQ channel activity in adult rats
Authors: Sedivy Vojtech , Hnilickova Olga , Smolkova Pavlina , Herget Jan 
Published: June 2021

Hypoxic stress at birth is linked with increased susceptibility to various diseases in adulthood. Perinatal hypoxia causes irreversible changes of the pulmonary vascular bed, some of which are more expressed in females. KCNQ (subgroup of voltage gated K+) channels have been suggested to play a role in pulmonary vascular reactivity. We hypothesized that perinatal hypoxia alters KCNQ channel activity in adulthood.Rats of the perinatal hypoxic group (PH) were born in a normobaric hypoxic chamber (FiO2 0.12; one week before the delivery) and kept in the chamber for one week after birth. Then they were raised in atmospheric air. Normoxic controls (N) were born and kept in room air. Both groups were divided according to gender and studied at the age of 20 weeks. We measured changes in perfusion pressure in isolated, saline‐perfused lungs. Dose‐response to a specific KCNQ channel inhibitor linopiridine (0.5–16 μM) was tested.In contrast to males, PH females have increased basal perfusion pressure and increased reactivity to K+ than N females. PH rats of both sexes are more sensitive to linopirdine. The increase in perfusion pressure in response to the highest linopiridine dose was 10.2 ± 1.6 mmHg in the PH group and only 5.0 ± 0.8 mmHg in the N group.Exposure to hypoxia in perinatal period increases KCNQ channel activity in adult rats.Supported by Cardiovascular Research Center MSMT 1M 0510 and Czech Science Foundation 305/08/0108.
Heat stress‐induced autophagic regulations in rat skeletal muscle
Authors: Yoshihara Toshinori , Naito Hisashi , Kakigi Ryo , Ichinoseki‐Sekine Noriko , Sugiura Takao , Katamoto Shizuo 
Published: June 2021

PURPOSETo examine the effects of acute heat stress on the regulation of autophagy in rat skeletal muscle.METHODSTwenty‐eight male Wistar rats (15 wk‐old) were used in this study. After 12h fasting, one leg was immersed in a hot water (43°C) for 30 min under anesthesia. The other leg was served as a control one. During the heat treatment, muscle temperature was recorded by a needle thermometer. The soleus muscle in both legs were removed before (Pre, n=4) and 0 (Post0, n=6), 30 (Post30, n=6), 60 (Post60, n=6) min or 24 hours (Post24h, n=6) after the heat stress. Phospho‐ and total‐Akt and mTOR, and LC3‐I/II protein expressions in the both muscles were determined by Western blotting technique.RESULTSMuscle temperature of the heat‐stressed leg was gradually increased to 41.8±0.3°C. In the soleus muscle, the levels of the autophagic makers LC3‐II were significantly increased in the Post60 group compared with Pre, Post30 and Post24h. Also, LC3‐I protein expressions were significantly decreased in the Post24h compared with Post0, Post30 and Post60. The Akt phosphorylation ratios of Post0 were significantly higher than any other groups. Moreover the phosphorylation level of mTOR was significantly increased in Post0 compared to Pre and Post24h.CONCLUSIONThese results show that acute heat stress lead to not only the activations of Akt/mTOR pathway but also the facilitation of protein turnover via autophagy.
Electrophysiological and molecular characterization of the two‐pore‐domain Potassium channels in murine fibroblasts of renal cortex
Authors: Kacik Michael , Kaistha Brajesh , Hoyer Joachim 
Published: June 2021

BackgroundPotassium channels have been proposed to play a key role in the progression of organ fibrosis. Recently, it has been shown that potassium channels control crucial fibroblast functions like proliferation and migration (Grgic., 2009).Our aims in this study are to characterize the role of new classes of potassium channels in murine renal fibroblasts (TFBs) and determine the regulation of the activity of these newly characterized potassium channels by different growth factors that are important in the pathophysiology of renal fibrosis.MethodChannel characterization were determined by whole cell patch‐clamp experiments of TFBs. In addition to channel function we tested the expression with real‐time‐PCR and the localization with immunofluorescence experiments.ResultsIn murine renal fibroblasts we detected the expression of six members of the two‐pore‐domain potassium channels (K2P) family. In the quantitative RT‐PCR experiments, TREK‐1, TREK‐2 and THIK‐1 were the most dominant arachidonic‐acid (AA) sensitive K2Ps.In whole cell experiments, we tested activators or inhibitors like changes in pH and temperature, mechanical stress, volatile anaesthetics, unsaturated fatty acids, metallic substances, antiarrhythmic and antidepressive agents. Our data suggest that THIK‐1 has a significant contribute to AA‐sensitive current.Mitogenic stimulation of fibroblast with profibrotic agents like b‐FGF and TGF‐β led to a downregulation of the channel activity in more than 70 %.ConclusionAside from previously described Ca2+‐activated potassium channels renal fibroblasts also possess a distinct array of K2P‐potassium channels. These channels are also regulated in the presence of profibrotic stimuli. In the clinical context K2P‐channels should be tested as pharmacological target in renal fibrosis.
Expressioneering™ Technology Accelerates Functional Expression and Crystallization of GPCRs for Drug Discovery
Authors: Sen Saurabh , Franz Laura , Mead David , Steinmetz Eric 
Published: June 2021

Structural biology of membrane proteins and GPCRs has seen major advances over the past few years, but overcoming the challenges of functional GPCR expression and structural stabilization are still time‐consuming and expensive. Our approach combines a powerful cloning strategy with novel transmembrane guides and co‐expressed helper proteins to significantly increase functional receptor expression and membrane insertion in E. coli.Lucigen's Expressioneering™ technology is a rapid and simple recombination based expression cloning method. We present here the use of this technology to optimize GPCR expression through fusion to a variety of transmembrane guides and novel visualization tags. We have been successful in obtaining conformationally active functional GPCRs with this system. We will also discuss unique new strategies for conformational stabilization of GPCRs through use of novel fusion partners and interacting proteins. Preliminary data using a novel fluorescent protein in the IL3 demonstrates retention of functional activity and may promote GPCR crystallization. Data obtained using Lucigen's Expresso CMV system are comparable with those obtained from the bacterial system.
Characterization of an anti‐pathogenic lactonase from Palk Bay metagenome
Authors: PANDIAN SHUNMUGIAH THEVAR KARUTHA , NITHYA CHARI , VISZWAPRIYA DHARMAPRAKASH 
Published: June 2021

ObjectivesScreening of metaclones for anti‐pathogenic activity and identification of anti‐pathogenic agents.MethodsScreening for anti‐pathogenic activity using Chromobacterium violaceum (ATCC 12472) and CV026 as indicators. Mode of action by TLC (Thin Layer Chromatography), HPLC (High Performance Liquid Chromatography) and PCR (Polymerase Chain Reaction).ResultsRecombinant fosmid clones (N=10200) were propagated in E. coli DH10B. When individually 1056 clones were used to challenge C. violaceum CV026, eight clones could able to inactivate the perception of QS signal HHL (C6‐HSL) by the Chromobacterium biosensor. Additional assays revealed that the metagenomic clone M8‐70 disrupted AHL signals, including C6‐HSL, heptanoyl‐ (C7‐ ), octanoyl‐ (C8‐) and decanoyl‐homoserine lactone (C10‐HSL), as well as the hydroxy‐ or oxo‐substituted derivatives at carbon 3, such as OC6‐HSL, OC8‐HSL and OHC8‐HSL. Culture supernatants of M8‐70 was examined for its effect on P. aeruginosa PAO1 (LasA, LasB, Pyocyanin, Pyoverdin and and Total protease), S. marcescens (prodigiosin production, swarming, secreted caseinase and lipase) QS system and distinct profiles were obtained for las and rhl QS genes. Furthermore, the growth curve analysis and cell count method made it clear that the supernatant of M8‐70 had no antibacterial activity toward P. aeruginosa PAO1 and S. marcescens. The results of the ring closure assay and the sequence analysis of the PCR products from M8‐70 (GenBank accession number HQ876560) revealed that its activity as lactonase activity.ConclusionIn this study a lactonase is reported through metagenomics approach from the sediment of Palk Bay which was found responsible for the inhibition of QS mediated virulence factors of Gram negative bacterial pathogens without inhibiting their growth.
Mediterranean Diet adherence is decreasing in healthy subjects over the years 1996–2010 also in Mediterranean area.
Authors: Trovato Guglielmo M , Martines G. F. , Catalano D. 
Published: June 2021

Mediterranean Diet score (range 0–55) was assessed in 8138 healthy non‐diabetic, overweight/obese subjects, 1996–2010, referred for US liver diagnostic and dietary counseling. Mediterranean food (Pasta and rice;whole‐grain bread, brown rice, legumes; Fruit; Green vegetables; Fish, poultry; Nonfat or low‐fat dairy products; olive oil) had assigned a score of 0: no consumption, a score of 1 for 1 to 4 times/w, 2 for 5 to 8 times/w, 3 for 9 to 11 times/w, 4 for 12 to 14 times/w, and 5 for more than 14 times/w; for westernized food (Red meat; Dairy products‐butter; Potatoes and eggs; Cakes) opposite scores were assigned (i.e., 0 when a participant reported more than 5 weekly consumption to: 5 for no weekly consumption, score 4 for 1 weekly consumption, 3 for 2 weekly consumption, 2 for 4 weekly consumption, 1 for 5 weekly consumption); wine and alcoholics (0–10 g of alcoholics from Red Wine for women score 5; 0–20 g of alcoholics from Red Wine for men score 5; each increment of 10 determines negative scores (20–30= −1. 30–40=−2. 40–50=−3. 50–60=−4, >60= −5 for men; 10 less for women and for all non‐wine alcoholics,: 10–20= −1; 20–30= −2; 30–40= −3; 40–50= −4; >50 = −5).There is a trend toward the loss of the adherence to Mediterranean Diet from 37.06±3.213 to 34.82±5.014 p<0,0001, not associated with BMI or physical activity change.Adherence to Mediterranean Diet is significantly yearly decreasing in Italy in healthy subjects.
Autoimmune disease (AID) in anorexia nervosa and bulimia nervosa
Authors: Gerhardt Ann 
Published: June 2021

We explored an impression, gained from a general health status questionnaire, that AID prevalence may be high in eating disorder patients. In the original study, 79 of 488 former eating disorder patients of an internal medicine practice, age ≥30 years at the time of the study, were found and 73 had completed a general health questionnaire with 30 questions related to 10 organ systems. No question mentioned any specific autoimmune disease. We contacted patients for more information about diagnoses.ResultsFifteen women (20.5%) had one or more AID diagnosis. These included Hashimoto's thyroiditis, celiac disease, systemic lupus erythematosis, rheumatoid arthritis, Crohn's disease, uncharacterized connective tissue disorder, combined variable immunodeficiency disease, interstitial cystitis, membranous glomerulonephritis and multiple sclerosis. AIDs were more common with advanced age (mean age 47.13± SD 9.27 vs. 41.40± SD 8.53, p=0.026). AID prevalence was significantly more common (34.8% vs. 14%, p=.041) in women who abused laxatives (N=23).ConclusionAIDs are common in this eating disorder population, and may be related to laxative abuse. The 20.5% prevalence is high compared to an estimated AID prevalence of 14% in all women ≥ 30 years of age, especially since none had Type I diabetes mellitus or psoriasis, the most common AIDs.Support from Sutter Institute for Medical Research
Lessons Learned in Conducting Nutrition‐Related Research in African American (AA) Older Adults
Authors: BABATUNDE OYINLOLA TOYIN , CAMPA ADRIANA , HIMBURG SUSAN P 
Published: June 2021

BackgroundTo reduce health disparity, the implementation of culturally sensitive strategies to improve participation of minority groups in nutrition research is needed. The objective of this report is to present lessons learned from outreaching and recruiting community‐based minority elders.MethodsParticipants were recruited through gatekeepers who were trusted by the target population and who belonged to the Churches and Senior Centers where African American (AA) elders routinely attend in three South Florida counties. Accessing and recruiting older AAs was challenging without these gatekeepers who acted as liaison between the church, group leaders, the participants and the investigator to negotiate agreement to participate in research. All data‐collecting forms were field‐tested to adapt them culturally. After 2 years, 130 participants were recruited, consented and participated in a six‐week education sessions on osteoporosis prevention.ResultsMean age of participants was 70.1 ±SD 10.4 y., 90% were women, and median education was 13 y. (range 8–20 y.). Attrition rate was 15.4%, 110 (84.6%) completed the program.Lessons LearnedAccessing AA older adults, an underserved and hard‐to‐reach group, requires determination and perseverance. Trusted gatekeepers, cultural adaptations and familiar social settings are needed to improve participation of minority groups in nutrition‐related research to reduce health disparity.
Genetic polymorphisms in AHR and CYP1A2 are associated with habitual caffeine consumption and are modified by age and smoking
Authors: Josse Andrea R , Costa Laura A Da , Campos Hannia , El-Sohemy Ahmed 
Published: June 2021

Recent genome‐wide association studies (GWAS) from populations of European decent have identified polymorphisms in AHR and CYP1A2 that are associated with caffeine/coffee intake. We examined whether these polymorphisms in AHR (rs6968865 and rs4410790) and CYP1A2 (rs2472297 and rs2470893) were associated with caffeine consumption in a Costa Rican population. Subjects (n=4,612) were participants from a case‐control study of gene‐diet interactions and myocardial infarction. Those with hypertension (n=1,692), or missing information on genotype (n=389), caffeine intake (n=4) or smoking status (n=4) were excluded. Sequenom MassARRAY was used for genotyping, and caffeine intake was assessed by a validated food frequency questionnaire. Compared to those consuming <100 mg/d of caffeine, subjects consuming >400 mg/d were more likely to be carriers of the T, C or T allele for rs6968865, rs4410790 and rs2472297, respectively. The corresponding odds ratios (OR [95% CI]) were 1.41 (1.03–1.93), 1.41 (1.04–1.92) and 1.55 (1.01–2.36). The multivariate‐adjusted ORs (95% CI) for rs6968865 were 1.44 (1.03–2.00) for all subjects, 1.75 (1.16–2.65) for non‐smokers and 1.15 (0.58–2.30) for smokers, and 2.42 (1.45–4.04) for >57 yrs and 1.00 (0.65–1.56) for ≤57 yrs. Our findings confirm previous results from GWAS linking polymorphisms in AHR and CYP1A2 with caffeine consumption, but only in non‐smokers and older adults.Grant Funding Source: Canadian Institutes of Health Research (CIHR)
Investigating piglet kidney vitamin A concentrations after high dose supplementation at birth
Authors: Gannon Bryan M , Arscott Sara A , Tanumihardjo Sherry A 
Published: June 2021

Potential metabolism and toxicity issues with infant vitamin A dosing procedures have led to a vested interest by the World Health Organization. Whole body retinoid trafficking was under investigation by determination of serum, liver, lung, kidney, adrenal, and spleen retinoid concentrations of piglets born from vitamin A depleted sows and dosed at birth with 25K, 50K, or 200K IU preformed vitamin A. These doses represent levels currently administered to newborn children and mothers in regions with high prevalence of vitamin A deficiency. Retinoids are known to be stored in these tissues with kidney being the primary extrahepatic storage site. In this study, kidneys were analyzed by grinding with sodium sulfate and adding C‐23 alcohol as an internal standard. Dichloromethane was used to extract retinol and retinyl esters. All samples were saponified, extracted, and injected onto an HPLC system to analyze total retinol content. A subgroup was also analyzed unsaponified to elucidate the retinyl ester profile. Data will be used collectively with other organ and serum data to have a complete model for vitamin A circulation and storage under a deficient status followed by high dose paradigm.Grant Funding Source: World Health Organization