bluuebunny/crossref_metadata_2025_split huggingface dataset:
Stressor and Multi‐Organ Antioxidant Profile following Early Life Maternal Deprivation in Albino Rats
Authors: Obafemi Faith Ayobamidele 
Published: April 2020

One‐week‐old albino pups were separated from their dams for a period of 24 to 48 hours. Sixty (60) rats were divided into 6 equal groups (n=10). Group A rats were separated for 24 hours and given glucose subcutaneously (200mg/kg) at 2, 7, 12 and 22 hours post maternal deprivation (MD). Group B rats received no intervention or treatment. Group C was subjected to 24 hours MD and given distilled water at regular intervals as group A. Group D rats had no MD but were given distilled water at same intervals as group A. Groups E and F were subjected to 48‐hour MD and administered Glucose or distilled water respectively at 2, 7, 17, 22, 27, 32, 42 and 47 hours post MD. Blood and tissue samples were collected for biochemical analyses.Significant weight gains were recorded in the groups with no maternal deprivation. The 48‐hour MD group supplemented with glucose competed favorably, showing a 7.2% weight gain over two days. At the other end of the spectrum, the 24‐hour MD supplemented with distilled water group recorded a 3% weight loss.The results show a graded rise in brain total protein values with time, with the highest values recorded in the 48‐hour MD + distilled water group and the lowest in the 24‐hour MD + distilled water group. Other organs showed no significant differences between groups. There were no differences across groups in all the organs assayed for nitric oxide (NO) either. For specific oxidative stress markers, Brain malondialdehyde (MDA) levels appear to be significantly lowest at the 48‐hour MD + distilled water group when compared with other groups. This correlated with a time dependent depletion in the brain antioxidant enzyme profiles studied. Brain GST, GPX and SOD all show highest concentrations in the 24‐hour MD + glucose group with lowest values recorded in the 48‐hour MD + distilled water group. Other organs did not show marked differences in between groups.Cortisol levels were assayed to evaluate stressor responses. The highest concentrations were seen in the 48‐hour MD + glucose group with the least seen in the 24‐hour MD + distilled water group. Consequently, parenteral administration of fluids (water and glucose) are a poor substitute for maternal nutrition although initial evidence points towards potential decline in brain antioxidant profile.Support or Funding InformationThe Tertiary Education Trust Fund (TETFUND) of Nigeria. The Faculty of Veterinary Medicine, University of Abuja, Nigeria.The Cardiorenal research group, Faculty of Veterinary Medicine, University of Ibadan; especially Professor Adeolu A. Adedapo, Dr. Tayo O. Omobowale, Dr. Ademola A. Oyagbemi and Dr. Jerry Afolabi.Brain Total Protein (TP) and Malondialdehyde (MDA) charts of all the groupsFigure 1
Role of protein kinase D1 in pulmonary hypertension in rats
Authors: Oku Shinya , Mizuno Yusuke , Sugawara Yoh , Goto Takahisa 
Published: April 2020

Protein kinase D (PKD) is known to be implicated in a variety of physiological functions and pathological conditions including cardiovascular diseases, such as cardiac hypertrophy and ischemia reperfusion injury. PKD has been thought to be a new target of the diseases for treatments. Previously, we showed that PKD played a role in pulmonary arterial contraction through MYPT1 phosphorylation and actin polymerization and that inhibition of PKD activity decreased right cardiac ventricular pressure in monocrotaline‐induced pulmonary hypertension (PH) model,. In this study, we further investigated a precise role of PKD in PH and in human pulmonary smooth muscle cells (HPSMCs).Material and methodsEight‐week‐old male rats were given either 60 mg/kg MCT or saline 4 weeks before experiments. Isometric tension was measured using pulmonary arterial ring samples to examine the effect of PKD inhibitor, CRT0066101 on norepinephrine (NE)‐induced contraction and phosphorylation of PKD, myosin phosphatase target subunit 1 (MYPT1), regulatory myosin light chain (RLC) and other PKD substrate proteins. Expression of PKD and phosphorylated‐PKD in rats lungs were also examined with immunohistochemistry using anti‐phospho‐PKD antibodies. We also investigated precise roles of PKD isoforms using HPSMCs with RNA interference.ResultsIn pulmonary arterial rings, basal expression of PKD1 and phosphorylated PKD were increased in pulmonary arteries in PH rats. NE increased phosphorylation of PKD, MYPT1 and RLC in ring samples during contraction, which were attenuated by pretreatment of CRT0066101. In immunohistochemistry, increased expression of PKD1 was observed in pulmonary arteries in PH rats (Fig
). In HASMCs, increased phosphorylation of MYPT1 and RLC with NE were significantly inhibited by PKD1 but not PKD2, 3 knock down.ConclusionsPKD1 may play role in contraction of pulmonary arteries and at least partially via MYPT1 phosphorylation. Increased PKD1 could be involved in PH development in rats.Support or Funding InformationGrant‐in‐Aid for Scientific Research (C) 17K11058Immunohistochemistry with anti‐PKD antibody in lung tissue from PH rats.Figure 1
Racial Differences in p66shc Activity in Activated Endothelial Cells
Authors: Aldokhayyil Maitha , Gomez Dulce H. , Kavazis Andreas N. , Brown Michael D. 
Published: April 2020

BackgroundOxidative stress plays a pivotal role in the development of endothelial dysfunction and the pathophysiology of atherosclerosis. P66shc is an adaptor protein that acts as a redox‐sensitive enzyme involved in mitochondrial reactive oxygen species (ROS) production. Upon endothelial cell activation, protein kinase C‐β2 (PKC‐β2) mediates Ser36 phosphorylation of p66shc, thereby allowing p66shc translocation from the cytosol to the mitochondria. Furthermore, we have shown that African American (AA) endothelial cells display higher levels of inflammation and oxidative stress markers compared to Caucasian American (CA) endothelial cells. Therefore, the purpose of this study was to investigate racial differences in p66shc activity in response to tumor necrosis factor‐α (TNF‐α)‐induced endothelial cell activation.MethodsHuman umbilical vein endothelial cells (HUVECs) from three AA donors and three CA donors were cultured and incubated with TNF‐α (10ng/ml, 4hr) to assess racial differences in p66shc, PKC‐β2 and endothelial nitric oxide synthase (eNOS) protein levels and bioactivity.ResultsEnzymatically active eNOS (p‐eNOSS1177) was significantly lower in AA HUVECs under both basal and TNF‐α incubation conditions (p= 0.01, Fig.
). Additionally, p‐Ser36 p66shc/total p66shc was significantly lower in AA HUVECs compared to CA HUVECs (p= 0.04, Fig. 3C). However, there was no group or treatment effect on any on the other variables measured.Conclusionp66shc represents an important molecular pathway that mediates ROS production and plays an integral role in the development of cardiovascular disease. According to our results, racial differences in endothelial function cannot be explained by p66shc. However, future research should focus on assessing cytosolic vs. mitochondrial p66shc levels in AA endothelial cells compared to CA.eNOS protein expression and bioactivity in CA and AA HUVECs after 4 hours of incubation with TNF‐α (10ng/ml).A) Densiometric quantification and representative Western blot of p‐eNOS. B) Densiometric quantification and representative Western blot of eNOS. C) p‐eNOS bioactivity. (* p= .01 vs. CA HUVECs, n= 4 independent experiments. Data are shown as mean ± SEM).Figure 1p66shc protein expression and bioactivity in CA and AA HUVECs after 4 hours of incubation with TNF‐α (10ng/ml).A) Densiometric quantification and representative Western blot of p‐p66shc. B) Densiometric quantification and representative Western blot of p66shc. C) p‐p66shc bioactivity. (* p= .04 vs. CA HUVECs, n= 4 independent experiments. Data are shown as mean ± SEM).Figure 2
Piezo‐electric based Ultraprecise/Ultratrace 3D Food Printing of Potato Resistant Starch
Authors: Oh Yourim , Lee Nam Keun , Rhee Jin-Kyu 
Published: April 2020

Recently, compared to traditional food production systems, 3D food printing has gained a lot of interest because of the potential benefits of being able to customized food products fabrication in colour, shape, texture, flavor and even nutrition. Successful printing of food objects in 3D food printing is critical, and high accuracy and precision food printing technology is required to reproduce delicate and complex food 3D structures. In 3D food printing, the properties of food materials, such as rheological properties, chemical properties and thermal stability, are significant for the structure accuracy and precision of printed objects. In this study, we tried to present a 3D structure layering method for potato resistant starch using drop‐on‐demand (DOD) printing method (piezoelectric system type) that can express the fine structure of food by inkjet printing technology. Nanocrystals cellulose (CNC) and sodium alginate (AS) were used to form a food 3D structure, and 3D printing optimal ink formulation ratio of CNC and AS (CNC:AS=80:20) was selected through preliminary experiments. The purpose of this study was to determine the possibility of 3D printing and printability after comparing rheological properties of mixtures with and without annealing by varying the content of potato resistant starch (PRS) in the mixture of CNC and AS. 3D food structure with various combination of potato resistant starch (RS) to a mixture of CNC and AS was printed, and compared the rheological properties of the mixture that had not been temperature‐treated and the one that had been annealed. After that, the optimal material combination for precise 3D printing was explored together with the physical properties of the printed object.Support or Funding InformationThis research was supported in part by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (2018R1D1A1B07045349), the High Value‐added Food Technology Development Program, the Ministry of Agriculture, Food and Rural Affairs (MAFRA), Republic of Korea (118059‐2) and the Ewha Womans University Research Grant of 2019.Piezo‐electric based Ultraprecise/Ultratrace 3D Food PrintingFigure 1
Treatment of Sleep Disordered Breathing With Leptin Loaded Exosomes
Authors: Pho Huy , Freire Carla , Kojima Ryo , Berger Slava , Ramsey Jacob D. , Batrakova Elena V. , Kabanov Alexander V. , Polotsky Vsevolod Y. 
Published: April 2020

RationaleSevere obesity leads to two types of sleep disordered breathing, including more prevalent obstructive sleep apnea (OSA), recurrent obstruction of the upper airway during sleep, and obesity hypoventilation syndrome (OHS) caused by impaired central nervous system responses to CO2. Adipokine leptin suppresses food intake, increases metabolic rate, stimulates breathing and ventilatory responses to CO2, and activates pharyngeal muscles improving upper airway patency during sleep. However, diet‐induced obesity (DIO) causes leptin resistance due to reduced permeability of the blood‐brain barrier (BBB) for leptin. Recent work with exosomes as nanocarriers for drug delivery vehicles have shown the possibility of overcoming the BBB to deliver therapeutic agents. We hypothesize that leptin loaded exosomes overcome BBB and treat OSA and OHS in DIO mice.MethodsMale DIO mice (n=6) were head mounted with EEG and EMG leads. Polysomnogram in the barometric plethysmography chamber was performed with IV administration of saline, free leptin, empty exosomes, and leptin loaded exosomes one week apart in a randomized cross‐over manner. Minute ventilation (VE) was measured through sleep/wake states and normalized to body weight. Upper airway obstruction was defined by the presence of inspiratory airflow limitation (IFL), as characterized by an early inspiratory plateau in airflow at a maximum level (VImax) while effort continued to increase.ResultsAs we have shown previously, DIO mice treated with placebo demonstrated upper airway obstruction, especially during REM sleep, and hypoventilation throughout both NREM and REM sleep. Leptin loaded exosomes nearly doubled VImax (p<0.05) and induced an approximately 1.8 increase in VE in REM IFL (p<0.05) compared to the IV saline, IV leptin, and empty exosome groups (Figure
), suggesting that upper airway obstruction was relieved. In addition, leptin loaded exosomes significantly increased VE during non‐flow limited breathing (p<0.05) compared to all other groups.ConclusionExosomal leptin delivery increased ventilation during both non‐flow limited breathing and flow limited breathing, suggesting a stimulatory effect on control of breathing and upper airway patency. Our results indicate that exosomal leptin can be considered for OSA and OHS pharmacotherapy in obesity.Leptin loaded exosomes presented increased ventilation in diet‐induced obese mice. Maximal inspiratory flow (VImax; mL/s). Minute ventilation normalized to body weight (nVE; mL/min/g).Figure 1
When is Muscle Sympathetic Nerve Activity ‘Abnormal’?
Authors: Badrov Mark B. , Keir Daniel A. , Tomlinson George , Notarius Catherine F. , Kimmerly Derek S. , Millar Philip J. , Shoemaker J Kevin , Floras John S. 
Published: April 2020

The sympathetic nervous system participates in both short‐ and long‐term cardiovascular regulation. The microneurographic technique, enabling in humans direct recordings from the populations of post‐ganglionic sympathetic neurons innervating the skeletal muscle vasculature (i.e. muscle sympathetic nerve activity; MSNA), has yielded key insights into mechanisms of sympathetic regulation of the circulation in health, and, importantly, its contributions to cardiovascular disease development and progression. Due to its invasive nature, as well as the technical challenges involved in site acquisition and burst quantification, its application has been limited primarily to trained investigators studying small, selected cohorts, rather than populations. In healthy individuals, MSNA burst frequency or burst incidence, as measured under supine resting conditions, exhibit considerable inter‐individual variability, yet are highly reproducible within individuals over time. However, such variability, as well as the limited data, has impaired the establishment of age‐ and sex‐dependent normative values. Therefore, we aimed to develop resting MSNA reference charts, representing MSNA percentile curves, in a large cohort of young and older men and women. Specifically, we retrospectively assembled a dataset of 654 healthy, unmedicated normotensive (<140/90 mmHg) individuals from four participating Canadian laboratories, including 396 men (aged 18–71 yrs) and 258 women (aged 18–81 yrs), in whom baseline resting MSNA was evaluated over a minimum of 5 min. Quantile regression was used to estimate the 5th, 20th, 50th (i.e. median), 80th, and 95th percentiles for resting MSNA burst frequency and burst incidence in men and women as a function of age. Quantiles were parameterized as functions of age by both a one‐ and two‐term fractional polynomial model with power selected from a ladder of values (i.e. −2, −1, −0.5, 0.5, 1, 2, 3). Next, an iterative fitting approach was applied to the MSNA versus age data to determine the fractional coefficients and the combination of values for the two powers from the ladder that provided the greatest goodness of fit (i.e. lowest χ2). The 5th, 20th, 50th, 80th, and 95th percentiles for MSNA burst frequency and burst incidence in men and women as a function of age are displayed in the figure
below. In conclusion, these MSNA percentile curves, developed from a large sample of men and women, provide age‐ and sex‐related reference values for resting MSNA levels in healthy, normotensive individuals, and, furthermore, by establishing 95% confidence limits, may help inform comparative studies of disease states referenced to age‐ and sex‐matched controls.Figure 1
Histone Deacetylase Inhibitors Enhance CRISPR‐Cas9 Cutting Efficiency
Authors: Bjornson Ymer M. , Huang Cody Y. , Castaneda Guadalupe , Yamomoto Emiko 
Published: April 2020

The CRISPR/Cas9 technology is a prominent genome‐editing tool as an adaptable nuclease capable of producing a double strand break at almost any genomic loci. However, the modification of hematopoietic stem and progenitor cells (HSPC) via the homology directed repair (HDR) pathway is still inefficient. We hypothesize that histone deacetylase inhibitors (HDACi), such as Valproic Acid (VPA) and Sodium Butyrate (NaB), could enhance HDR efficiency by increasing the accessibility of the genome‐editing machinery. To address the potential utilization of HDACi therapeutically, we began by assessing the effect of VPA and NaB on cell growth and viability. No statistically significant effect on cell growth or viability was observed at concentrations as high as 50mM in a myeloid‐erythroid leukemic cell line (K562 cells). However, K562 cells only represent one lineage of the HSPC progeny. Therefore, we also treated an immortalized T lymphocyte cell line (Jurkat cells), representative of the lymphoid lineage, with either VPA or NaB. No effect on the growth pattern or viability of Jurkat cells was evident at concentrations as high as 5mM VPA and NaB. However, Jurkat cells did appear to be more susceptible to HDACi. Therefore, we ascertained that concentrations of HDACi lower than 5mM would be tolerated in HSPCs. At concentrations as low as 0.0005mM HDACi an enhancement in CRISPR cutting efficiency was evidenced as assessed in both K562 cells and Jurkat cells. This enhancement did not appear to be locus specific, in that two sets of CRISPR guide RNAs were analyzed revealing similar trends of enhancement. If the same trend follows, treatment with HDACi can be expanded to virtually any genome editing experiment utilizing the CRISPR‐Cas9 system to enhance cutting and thus therapeutic gene editing.Support or Funding InformationCSUPERBEffect of Valproic Acid and Butyrate on the Growth Curve of Hematopoietic Cell Lines A) Schematic representation of the experimental design utilized to test the effect of HDACi on the division of two hematopoietic cell lines. B) K562 cells treated with 300mM and 100mM VPA were not observed to divide. C) K562 cells treated with 500mM NaB were not observed to divide. D) Jurkat cells appear to be more susceptible to VPA. E) Jurkat cells treated with 50mM NaB proliferated at a much slower rate.Figure 1
Elucidate Glycosyltransferase Specificities And Interactions for Rational Glycoengineering
Authors: Liang Chenguang , Chiang Austin W.T. , Hansen Anders H. , Arnsdorf Johnny , Schoffelen Sanne , Sorrentino James T. , Kellman Benjamin P. , Bao Bokan , Voldborg Bjørn G. , Lewis Nathan E. 
Published: April 2020

The emergent market of new therapeutic glycoprofiles demands cost‐and‐labor efficient N‐linked glycoengineering, such as rational glycoengineering. However, rational glycoengineering remains challenging due to our limited understanding of the complex glycosyltransferase (GT) interactions. Predicting glycosylation outcomes of GT knockins/knockouts requires comprehensive and context‐specific knowledge of GT activities. To combat this issue, we proposed an improved low‐parameter Markov chain model to learn GT activities and used the learnt information to predict glycoprofiles impacted by complex glycoengineering. Specifically, we have quantified the impact of GT isozyme specificities and interactions by fitting the model with a compilation of single‐GT‐knockout erythropoietin (EPO) glycoprofiles for CHO cells. We then showed that the quantified GT impact can be generalized by our model framework to predict complex multiple‐GT‐knockout glycoprofiles of four diverse therapeutic drugs, including EPO, Rituximab, Enbrel, and alpha‐1 antitrypsin. Therefore, the proposed methodology allowed us to systematically unravel the complicated GT interactions beyond simple enzyme rules and with little a priori information. Such approach will further promote progress in rational glycoengineering by broadening our scope on the genetic basis of glycosylation, expediting product development for biopharmaceutical research.Support or Funding InformationThis work was conducted with support from the Novo Nordisk Foundation provided to the Center for Biosustainability at the Technical University of Denmark (NNF10CC1016517: A.L., A.W.T.C., A.H.H., B.G.V.) and NIGMS (R35 GM119850: N.E.L.)Figure 1
A Universal CAR‐NK Cell Targeting Various Epitopes of HIV‐1 gp160
Authors: Lim Rebecca Marie , Rong Liang , Xie Jianming 
Published: April 2020

Just like cancer cells that express cell‐surface tumor antigens, HIV‐infected cells express the envelope glycoprotein gp160 upon viral budding. Hence, engineering T cells and natural killer (NK) cells with anti‐gp160 chimeric antigen receptors (CAR) has emerged as a promising strategy to eradicate HIV‐infected cells. However, current CARs are limited by targeting a single epitope on gp160, which cannot counter the enormous HIV diversity. Here, we report the development of a universal CAR‐NK cell, which recognizes 2,4‐dinitrophenyl (DNP). This anti‐DNP CAR‐NK cell can be redirected to target various epitopes of the HIV envelope using DNP‐conjugated anti‐gp160 antibodies as adaptor molecules. We have shown that this CAR‐NK cell can recognize and kill mimic HIV‐infected cell lines expressing HIV‐1 subtypes B and C gp160. We additionally found that anti‐gp160 antibodies targeting membrane‐distal epitopes were more likely to activate universal CAR‐NK cells compared with those targeting membrane‐proximal epitopes. Moreover, we demonstrated that the universal CAR‐NK cells exhibited similar efficacy to the conventional CAR‐NK cells in targeting gp160‐expressing cells. Given that numerous anti‐gp160 antibodies are readily available, this universal CAR‐NK cell can potentially be used to overcome HIV diversity and to eradicate HIV‐infected cells.Support or Funding InformationThis study is supported by a startup fund from University of Southern California School of Pharmacy (J.X.) and a Pre‐Doctoral Fellowship in Pharmaceutical Sciences from the American Foundation for Pharmaceutical Education (R.M.L.).Our proof‐of‐concept study demonstrates that we can direct universal CAR‐NK cells with DNP‐labeled broadly neutralizing antibodies to target and kill multiple variants of HIV‐infected cells. This strategy can help overcome HIV‐1 diversity and enhance eradication of infected cells, which is essential to finding an ultimate universal cure to HIV.Figure 1
MicroRNA‐93, a Novel Potential Non‐Invasive Biomarker for Polycystic Ovary Syndrome (PCOS) Diagnosis
Authors: Chen Elizabeth , Wu Hsioa Li , Chen Yen Hao 
Published: April 2020

PCOS is one of the most common endocrine disorders and affects at least 7–9% of reproductive‐aged women. However, PCOS diagnosis is a challenge for physicians because of the constellation of symptoms. From our previous studies, we found that the expression of microRNA‐93 (miR‐93) was up‐regulated in adipose tissue and related to the high incident of insulin resistance in PCOS women. As a result, we generated an adipose‐selective miR‐93 overexpression mouse (miR‐93AT+ mice) model which shows a PCOS‐like phenotype by using Cre/loxP DNA recombination. The level of miR‐93 expression in tissues was investigated by the q‐PCR method. The expression of miR‐93 was specifically increased in adipose tissue but not in muscle, liver, heart, or kidney tissues in miR‐93AT+ mice. However, the level of miR‐93 was overexpressed in the blood of the miR‐93AT+ mice, similar to results found in PCOS women. In addition, the level of miR‐93 was overexpressed (2.5 fold) in the feces of miR‐93AT+ mice. These data suggest that fecal miR‐93 may be applied as a potential novel non‐invasive biomarker for the diagnosis of PCOS. Additional studies of miR‐93 expression in feces from PCOS women are required to confirm this hypothesis.Figure 1