bluuebunny/crossref_metadata_2025 huggingface dataset:
Sub‐neurodegenerative Levels of Binge Alcohol Reduce Calcium‐independent Phospholipase A2 (iPLA2) Levels in Rat Brain: An Unrecognized Initial Sign of Neuroinflammation
Authors: Schreiber Jennifer Ann , Tajuddin Nuzhath , Kouzoukas Dimitrios , Kim Hee‐Yong , Collins Michael
Published: April 2020

Selective degeneration of dentate granule cells in hippocampus (HC), pyramidal cells in entorhinal cortex (EC), and olfactory bulb neurons occurs in adult male rats subjected to severe alcohol (ethanol) binges over 4 days that generate maximal blood alcohol levels of 350–450 mg/dl. Concurrent with brain damage specifically in these brain regions, significant alterations are elicited by alcohol in the levels of phospholipase A2 (PLA2) enzymes, i.e., (1) increases in Ca2+‐dependent cytosolic cPLA2 and secreted sPLA2, two membrane phospholipid‐metabolizing enzymes causing neuroinflammation via mobilization of omega‐6 arachidonic acid, a largely pro‐oxidative neurocellular mediator; and (2) reductions in Ca2+‐independent iPLA2, known to selectively regulate membrane release and turnover of anti‐inflammatory, anti‐oxidative omega‐3 docosahexaenoic acid (DHA). Furthermore, these neurodegenerative and neuroinflammatory changes are replicated in vitro, along with a loss in DHA content itself, in rat organotypic adult‐age HC‐EC slice cultures binge‐treated with 100 mM (~460 mg/dl) alcohol. Importantly, DHA pre‐supplementation of the slices prevents the neurodegeneration and PLA2 changes caused by alcohol (Tajuddin et al. 2014). The objective of this study was to determine if phospholipid‐mediated neuroinflammatory changes exist from alcohol in the absence of neurodegeneration. Here we report that rats subjected to less severe binges ‐ and thus lower blood alcohol levels of ~200 mg/dl, which are insufficient to cause detectable brain damage and/or neuroinflammatory cPLA2 and sPLA2 elevations ‐nevertheless cause significantly reduced iPLA2 levels that are largely reversed by DHA supplementation. The results imply that iPLA2 depletion and, we suspect, loss of cellular/membrane DHA (that iPLA2 regulates), are critical, previously unappreciated first indicators or markers of brain neuroinflammation due to binge alcohol. Similar changes in iPLA2, potentially due to its sensitivity, may have a bearing on other forms of brain insults causing neuroinflammation and ultimately neurodamage.

Support or Funding Information

Research support: NIH U01 AA018279 (MAC and HYK) and NIH T32 AA013527 (Loyola Medical Center Alcohol Research Program)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Hepatocytes Uptake Clearance of Organic Anion Transporting Polypeptide (OATPs) Substrates in Human and Cynomolgus Cryopreserved Hepatocytes
Authors: Liang Xiaomin , Deforest Natalie , Park Yeojin , Hao Jia , Zhao Xiaofeng , Lai Yurong
Published: April 2020

Transporter‐mediated clearance plays significant roles in drug bioavailability and elimination, and evaluation of potential transporter‐mediated drug‐drug interactions (DDI) is recommended by regulatory agencies during drug development. Currently, the direct extrapolation from in vitro to in vivo is a common approach for human clearance prediction for the compounds that are metabolized by hepatic cytochrome P450 enzymes; however, challenge remains when using the similar approaches in prediction of transporter‐mediated clearance. Recent publications attempted to establish the in‐vitro‐to‐ in vivo scaling based on hepatocyte uptake studies and in vivo pharmacokinetic (PK) data from the preclinical animals. However, the differences in homology of transporter proteins and the associated substrate specificity in transporters between human and preclinical animals complicate the prediction using the correcting factors obtained from the preclinical animals' in‐vitro‐to‐ in vivo scaling. In this study, we obtained the in vitro hepatic uptake clearance in cynomolgus monkey and human for 14 literature compounds that were known as OATPs substrates and had clinical DDI reported. Hepatic uptake was conducted using the oil‐spin method. For each selected substrate, the uptake assay was conducted in the time course from 0.25 minute to 15 minutes in KHB buffer with 10% serum. The uptake clearance was estimated from the linear uptake phase. For each selected substrate, the uptake clearance was characterized in three lots (including single donor and pooled donors lots) of human and monkey cryopreserved hepatocytes. In addition, we tested different lots of human (7 lots) and monkey (10 lots) cryopreserved hepatocytes using rosuvastatin as a control probe to examine the variability among lots and donors. In summary, rosuvastatin uptake clearance in cynomolgus monkey hepatocytes was about 2.5 folds higher on average compared to that in human hepatocytes. Generally, for other OATPs substrates studied, hepatic uptake clearance in cynomolgus monkey appeared to be higher than that in human. Furthermore, variability of uptake clearance was also observed among lots and donors. The results obtained in the current investigations are useful for human prediction through understanding the source of variations and establishing the correlations of in vitro‐in vivo scaling.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular characterization of eel Lutropin/Chorionic Gonadotropin Receptor mutants
Authors: Byambaragchaa Munkhzaya , Min Kwan‐Sik
Published: April 2020

Whereas the ligand mutations that have been found to represent usually loss‐of‐function mutations, the G protein coupled receptor mutations can be both of the gain‐of function and loss‐of function type. There are a lot of number naturally occurring mutations of the lutropin/chorionic gonadotropin receptor (LH/CGR) gene associated with reproductive disorders. Although the hormone binding and activation properties of these mutants are generally well characterized in hLHR, little is known about their regulation and signal transduction if it occurs in eel LH/CGR. With this in mind we examined the effects of the several constitutive activation and inactivation mutations which are occurred in highly conserved residues of LH/CGR such as: activation mutations (M410T at TM2; L469R at TM3; D590Y at TM6) and inactivation mutations (D383N at TM2; Y546F at TM5) in eelLH/CGR consisting of 705 amino acid.

To assess the functional effects of 5 receptor mutations directly, wild‐type (WT) and mutant receptors were transiently expressed in CHO‐K1 cells. We evaluated the basal and cAMP stimulation by recombinant eel Luteinizing hormone (rec‐eelLH). The constitutive activation mutants all had elevated levels of basal cAMP. But inactivation mutants were remarkably decreased cAMP production in response to rec‐eelLH. Two of activation mutations (L469R and D590Y) apparently activated eelLH/CGR with the 19.8 and 8 fold elevated basal cAMP level. However M410T mutation showed only 4 fold elevated basal cAMP level, it could be possible to activate eelLH/CGR in a weak manner. Maximal responses of activation mutants were 7.2–46.7% lower than that of WT. The EC50 in two inactivation mutations significantly retarded the cAMP accumulation to 42.3 ng/ml and 1181 ng/ml, respectively.

Support or Funding Information

This work was financially supported by the Korean Research Foundation Program (2018R1A2B6007794)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The myeloid lineage‐determining transcription factor PU.1 induces enhancer‐promoter looping that promotes IL‐1β enhancer and messenger RNA production in non‐myeloid melanoma cells
Authors: Kim Sung Ouk , Cho William , Ha Soon‐Duck
Published: April 2020

The DNA‐binding protein purine rich (PU.1) is a lineage determining transcription factor required for development of myeloid cells such as monocytes/macrophages. PU.1 is regarded as a pioneering transcription factor for macrophage differentiation due to its ability to bind “closed” genomic sites and initiate/maintain “open” chromatin state in macrophage‐restricted response genes. To date, however, precise mechanism of PU.1 in remodeling chromatin is yet to be elucidated. The non‐myeloid melanoma B16.BL6 cells response to the bacterial cell wall component lipopolysaccharide (LPS) and activate the transcription factor NF‐κB, but do not produce cytokines. Ectopic expression of PU.1 rendered these cells express the cytokine interleukin 1‐β (IL1b) in response to LPS. PU.1 induced chromatin remodeling that caused interaction between the IL1b promoter and enhancer (located ~10 kb upstream of the IL1b transcription start site) in non‐activated cells; upon activation, PU.1 recruited NF‐κB to these genomic regions. The N‐terminal domain (1–30) of PU.1 was important for both promoter‐enhancer looping and IL1b eRNA/mRNA production; whereas, the acidic and glutamine rich domains (33–100) were required for eRNA/mRNA production but dispensable for the DNA looping. eRNA production and histone acetylation were required for NF‐κB recruitment and IL1b mRNA expression, but not for the DNA looping. Altogether, this study indicates that PU.1 induces enhancer‐promoter looping that sequentially allows activation‐dependent histone acetylation, recruitment of NF‐κB and production of enhancer RNAs, all of which are required for an optimal IL1b production.

Support or Funding Information

The Natural Sciences and Engineering Research Council of Canada ‐ Discovery Grant (RGPIN‐2018‐05514)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis and the identification of DNA binding sites for the cI repressor of Bacteriophage ɛ34
Authors: McGee Lauren , Gildea Logan , Goodson Richardria , Villafane Robert , Jackson Doba , Brooks Ryan
Published: April 2020

Gene 46 of ɛ34 encodes a protein that is similar to other phage repressors. It has a C‐terminal protease domain which is about 80% identical to the repressors of phage λ, but its DNA binding N‐terminal portion is only about 18% identical to its closest known relatives in phages λ, and Lex A. The N‐terminal DNA‐binding domain is homologous to the HTH_XRE family of DNA‐binding domains. In another study we have generated homology models of this proteins and its interaction with DNA [1].

The main goal of this project was to initiate a structure‐function study on the cI repressor protein of phage ɛ34. The cI repressor of ɛ34 protein with a 6X polyhistidine tag was first isolated and purified to homogeneity. We have identified regions for cI binding by using bioinformatics and gel‐mobility shift assay using PCR fragments encompassing the intergenic region of the cI and cro gene. Our binding sites compare well with the known intergenic regions of other phages such as λ, Φ11, 434. This project was done as a collaborative venture between two laboratories, Huntingdon College (Dr. Doba Jackson) and Alabama State University (laboratory of Dr. Robert Villafane). This project is in line with Dr. Robert Villafane's research into the Genomic analysis of bacteriophage ɛ34 of Salmonella enteric serovar Anatum.

Support or Funding Information

Support Support for this work was provided by Alabama State University and Huntingdon College.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Novel Rigid Glycomimetics to Inhibit Influenza Infection
Authors: Woods Robert J. , Ji Ye , Hendel Jenifer , Smith Hannah , Glushka John , Wu Nicholas , Wilson Ian A. , Tompkins S. Mark , Murphy Paul
Published: April 2020

Small‐molecule anti‐influenza agents, such as zanamivir and oseltamivir are neuraminidase (NA) inhibitors limit the release of progeny virions from infected cells through competitive binding to the viral NA. However, these drugs do not block infection. Vaccination is the most common approach for preventing influenza infection, but vaccine creation takes approximately 6 months and requires prior knowledge of the circulating virus strains. The impact of this limitation was evident in the 2009 swine flu pandemic, against which the seasonal vaccine was completely ineffective. The urgent need to develop complementary strategies to vaccination or NA inhibition is emphasized by the fact that between 2006 and 2009 seasonal flu developed almost complete resistance to oseltamivir, resulting from a single spontaneous mutation in the viral NA.

As a potential alternative to current flu antivirals, we are developing glycomimetics that block the interaction between the viral hemagglutinin (HA) and the host glycans that serve as flu‐receptors. This interaction is essential for viral adhesion, and the universal receptor for flu A [15] and B [16] viruses is Sia‐Gal (sialic acid, Sia, a.k.a. neuraminic acid, linked to galactose, Gal). HAs from either human (H1N1) or avian‐infective (N5N1) flu strains bind to di‐ or trisaccharides terminating in Sia‐Gal regardless of whether the Sia‐Gal linkage is α2,6 or α2,3. This observation suggested that a small Sia‐Gal glycomimetic might be developed that could inhibit a broad range of flu A strains, including flu B, and was critical in the design of our glycomimetic framework (FB127).

Here were report that FB127 shows dose‐dependent inhibition of the infection of MDCK cells by a range of flu viruses from group 1 (H1N1, H5N1), group 2 (H3N2), and flu B, and that the influenza neutralizing ability of FB127 is within the same range as reported for glycodendrimers. Because FB127 is a synthetic small molecule, it can be rationally modified, and manufactured with a high degree of quality control and reproducibility. In this regard, FB127 is amenable to a number of formulations, including aerosol, oral, and intravenous routes that provide great flexibility in drug delivery.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dysfunction of a microglia subtype produces sex‐linked anxiety
Authors: Traenkner Dimitri
Published: April 2020

Microglia are immune cells of the central nervous system. Different types of microglia exist, yet we incompletely understand to what extend subtypes serve different functions. To test this, we used a selective genetic ablation strategy in mice. Intriguingly, loss of one microglia subset causes severe anxiety exclusively in females. Moreover, this behavior develops at the time of sexual maturity, suggesting an important role of female sex hormones in this anxiety model. Our findings suggest a mechanistic link between sex and genetics, two major risk factors for anxiety disorders in people.

Support or Funding Information

National Institute of Health grant R01MH093595

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Intralipid Infusion in Pregnant Rats Induces Plasma Angiogenic Imbalance, Inflammation, and Intrauterine Growth Restriction
Authors: Duncan Jeremy W. , Bergeron Matthew , Bradshaw Jessica L. , Spradley Frank T. , Granger Joey P.
Published: April 2020

Preeclampsia is a pregnancy specific disorder characterized by maternal hypertension and proteinuria and is associated with low birth weight, vascular dysfunction, and an increase in inflammatory and anti‐angiogenic factors. Recent studies have shown that obese women are 4 times more likely to develop preeclampsia. In addition, preeclamptic patients have higher levels of circulating free fatty acids (FFA) and other lipids compared to normal pregnant women. While a high fat diet can lead to reduced uteroplacental perfusion, vascular and placental dysfunction, the mechanism whereby lipids contribute to these outcomes in preeclampsia is unknown. To test the hypothesis that lipid administration induces placental and vascular dysfunction, pregnant rats were infused intravenously with 20% Intralipids (50–100 μL/kg/min) over a 2 h period on gestational days (GD) 11–18 or ‐19. On GD 19, mean arterial pressure (MAP), pup weight, placental weight, blood and tissues were collected. Third order mesentery arteries and aortas were used for vascular reactivity studies, and placental villi were cultured for 48 h to measure the production of angiogenic factors ex vivo. Infusion of intralipids resulted in no change in MAP, vascular reactivity, or placental villous angiogenic factors. However, infusion of intralipids in pregnant rats caused a significant increase in placental weight (0.66 ± 0.01 g vs 0.61 ± 0.01 g; p<0.01) and fetal absorption (13.8 ± 5.9% vs 0.85 ± 0.85%; p<0.05), and a decrease in pup weight (2.37 ± 0.02 g vs 2.57 ± 0.02 g; p<0.01) compared to control rats. Furthermore, plasma tumor necrosis factor‐alpha (TNF‐α; 24.82 ± 6.21 pg/mL vs 12.24 ± 1.52 pg/mL; p<0.05) and soluble fms‐like tyrosine kinase (sFLT‐1; 20.08 ± 5.76 pg/mL vs 4.42 ± 2.04 pg/mL; p<0.05) were increased, while plasma placental growth factor (PlGF; 28.57 ± 5.65 pg/mL vs 51.19 ± 8.82 pg/mL; p<0.05) was decreased in intralipid‐infused pregnant rats compared to control rats. These data indicate that increasing lipid intake during pregnancy induces intrauterine growth restriction coupled with a pro‐inflammatory and anti‐angiogenic state. Further studies to determine the factors associated with intralipid‐induced fetal outcomes are warranted and could provide a better understanding of the role of elevated lipids/FFA as a risk factor for preeclampsia.

Support or Funding Information

P01HL051971, P20GM104357, and T32HL105324.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Non‐Small Cell Lung Carcinoma Conditioned Media on Skeletal Muscle Cell Proliferation and Metabolism
Authors: Stott Nicole L , Beach Cassandra , Marino Joseph S
Published: April 2020

Lung cancer is the second most common cancer and one of the largest causes of cancer‐related mortality worldwide. 80–85% of all new lung cancer diagnoses are Non‐Small Cell Lung Carcinoma (NSCLC), suggesting a great need to continue elucidating the metabolic mechanisms surrounding this prevalent disease. Due to the oncogenic environment surrounding NSCLC growth, factors released into the environment could have a direct impact on other tissues. In particular, skeletal muscle, a leading insulin‐sensitive and metabolically active tissue, is subject to wasting in many cancer patients. The purpose of this study was to determine how NSCLC conditioned media affects C2C12 skeletal muscle cells proliferation and metabolism. C2C12 mouse skeletal muscle cells were grown for 24 hours and serum‐starved overnight with serum‐free media to induce quiescence. Cells were treated with equal volumes of either fresh growth media or conditioned media from proliferating NSCLC cells. C2C12 myoblast proliferation was assessed every 24 hours for 3 days via an MTT assay using 3‐(4,5‐Dimethyltiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide. Proliferating C2C12 myoblasts treated with fresh growth media or conditioned media from NSCLC cells showed significant proliferation (p=0.0016) over 72 hours. Treatment with NSCLC conditioned media did not induce any significant increases in myoblast proliferation when compared to control C2C12 cells. In conclusion, these data suggest that C2C12 myoblasts did not respond adversely to NSCLC conditioned media. Experiments are ongoing to investigate alterations in metabolic and myogenic signaling pathways, as well as extended time courses.

Support or Funding Information

This work was supported by the Targeted Research Internal Seed Program, JSM.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Efficiency vs. Competency Development: What Interprofessional Health Professions Students Learn From, With, and About Each Other During Anatomy Dissection
Authors: Nelsen Sylvia , Tucker Constance
Published: April 2020

The gold standard for anatomy instruction has traditionally involved a combination of lecture‐based learning and hands‐on discovery through cadaveric dissection. However, a full dissection is time‐intensive, and recent curricular changes are challenging educators to deliver content via more time‐efficient modalities. As a result, laboratory hours are often a place where reductions in instructional time are made. This tension between time and meaningful activity is heightened as curricular innovations such as simulation, 3D visualization, and prosection are shown to reduced teaching time and increase the student: cadaver ratios. However, the dissection experience is not only important for helping students understand the structure and function of the human body. It is also instrumental in helping students develop skills and attitudes related to professionalism such as teamwork, communication, and respect for each other and their future patients, learning opportunities that are often referred to as the “hidden” curriculum (Escobar‐Poni & Poni, 2006; Warner & Rizzolo, 2006). More recently, the ability of healthcare providers to work effectively in interprofessional teams has been added to this list of skills, and dissection experiences have been designed to explicitly incorporate interprofessional experiences into anatomy lab curricula (e.g., Fernandes et al, 2015). To date, the impact of an interprofessional learning environment within the anatomy lab on learners' perceptions of teamwork has not been thoroughly examined. To explore the impact of the interprofessional learning experience, students were asked to write a short essay reflecting on what they learned from their teammates and the donors, and how that will affect the way they approach clinical care and working in professional teams. Constant comparative analysis was used to compare themes from essays written by dental students working in interprofessional teams comprised of dental, physician assistant, and radiation therapy students to those written by dental students working in teams segregated by profession. Results of the study highlight the unique contributions of an interprofessional dissection environment on the learning experience of health professions students.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mediation of Vesicular Fusion by SNARE Proteins during Macroautophagy
Authors: Shannon Daniel , Glass Jen , Bartlett Matt , Bissmeyer Teddy , Bradley Mariah , Buehler Sam , Campman Scotty , Glass Jake , Gutzwiller Leah , Hoffman Trevor , Kellogg Nick , Mahan Erin , Peters Kyle , Poetker Garrett , Schaller Stephanie , Diao JaiJie
Published: April 2020

The Moeller SMART Team in conjunction with Dr. JiaJie Diao at the University of Cincinnati Medical College and MSOE Center for Biomolecular Modeling used 3‐D modeling and printing technology to study the role of Soluble NSF Attachment Protein Receptor (SNARE) proteins in macroautophagy, a process that recycles damaged organelles, proteins, and microbes in the cell. It begins with the formation and maturation of an autophagosome. The process ends with the fusion of a fully matured autophagosome to a lysosome membrane, which is mediated by SNARE proteins. Vesicular fusion is initiated by the formation of a tetrameric alpha‐helix complex that is formed through the binding of three helical SNARE proteins. One of these SNARE proteins, VAMP8, is located on lysosomes and the other two, syntaxin17 and SNAP‐29, are located on the membrane of autophagosomes. The helices of these proteins coil tightly in the alpha‐helix complex which provides the energy for fusion. To reset the conformation of SNARE proteins, N‐Ethylmaleimide‐Sensitive Factor (NSF) proteins use ATP and water to recycle SNAREs. Alpha‐SNAP, an ATPase, which helps to facilitate this process, returns SNARE to its original high energy state. If fusion is unable to occur in autophagy, the cell accumulates autophagosomes, causing lysosome storage diseases such as Parkinson's and Alzheimer's disease. By 3‐D printing the coiled alpha‐helix complex, our SMART team intends to describe this crucial element of membrane fusion and the disorders associated with macroautophagy.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Total Sleep Deprivation Does Not Adversely Affect Arterial Stiffness, Wave Reflection and Aortic Pressure in Young Healthy Men
Authors: Lapierre Stephanie , Holmer Brady J , Lim Jisok , Sakarya Yasemin , New William , Eagle Michael , Hwang Chueh‐Lung , Schwartz Cody , Bhattal Gurjaspreet , Maharrey Phillip , Handberg Eileen M , Christou Demetra D , Alviar Carlos L
Published: April 2020

Sleep deprivation is a common problem and may contribute to increased risk for development of cardiovascular disease. The influence of sleep deprivation on arterial stiffness, wave reflection and aortic pressure, independent predictors of cardiovascular disease, has not been fully elucidated. Therefore, the purpose of this study was to investigate the acute effects of total sleep deprivation (TSD) on arterial stiffness, wave reflection and aortic pressure in young healthy men. Twelve participants (22 ± 1 yrs, means ± SE) completed this study. Participants were free of clinical disease and sleep disorders, and did not use tobacco products, or take medications. Arterial stiffness (carotid to femoral pulse wave velocity; cfPWV), wave reflection (aortic augmentation index; AIx), and aortic blood pressures (systolic and diastolic blood pressure; aoSBP and aoDBP, and pulse pressure aoPP) were assessed using a validated cuff‐based device (SphygmoCor XCEL, AtCor Medical). Measures were obtained at the same time in the morning following a night of regular sleep (CON) and a night of TSD. During TSD, participants rested quietly under supervision and abstained from caffeine for ≥20 hrs. The average duration of sleep deprivation was 26.3 ± 1.3 hrs. In response to TSD, cfPWV did not significantly change (6.6 ± 0.2 vs. 6.4 ± 0.2 m·s−1; CON vs. TSD; P = 0.1). Aortic pressures (aoSBP: 104 ± 2 vs. 105 ± 2 mmHg; aoDBP: 69 ± 2 vs. 71 ± 3 mmHg; and aoPP: 34 ± 2 vs. 34 ± 1 mmHg; P≥0.5) and AIx also remained unchanged in response to TSD (9 ± 3 vs. 5 ± 2 %; P = 0.1). In conclusion, TSD does not adversely affect arterial stiffness, wave reflection or aortic pressure in young healthy men. Future investigations should explore the influence of TSD in populations at high risk for developing cardiovascular disease.

Support or Funding Information

This work was supported by the Gatorade Trust through funds distributed by the University of Florida, Department of Medicine.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial Estrogen Receptor α/β Protein Ratio is Associated with Circulating Estrogen Levels in Healthy Women
Authors: Park Young‐Min , Keller Amy C , Runchey Shauna S , Brubaker Ashley , Hildreth Kerry L , Moreau Kerrie L
Published: April 2020

OBJECTIVE

Vascular aging, featuring endothelial dysfunction, precedes an increased risk of cardiovascular disease (CVD). The menopause transition (i.e., perimenopause) is characterized by changes in ovarian hormone and vascular health. We previously reported a decline in endothelial function that was correlated with lower estradiol (E2) and estrone levels. Biological effects of E2 on endothelial function are mediated via estrogen receptors (ER). We determined whether ER α and β activation (i.e. nuclear translocation) and the ER α/β ratio in endothelial cell (EC)s differed by menopause stages and whether they were associated with reduced endothelial function across menopause stages.

METHODS

EC samples were harvested from 67 healthy, fasted women (aged 30–70 years) classified as premenopausal (Pre; regular menstrual cycles; n=15, 33±6yrs; mean±SD), early perimenopausal (EPeri; > 2 cycles with cycle length changes of ≥7 days; n=15, 49±3yrs), late perimenopausal (LPeri; ≥2 but <12 months of amenorrhea; n=17, 49±4yrs), or postmenopausal (Post; n=20, 56±5yrs). ECs were analyzed for whole cell and nuclear ER α and β protein content by immunofluorescence. Endothelial‐dependent vasodilation was measured by brachial artery flow‐mediated dilation (FMD) using ultrasound.

RESULTS

ERα nuclear/cell protein ratio (a surrogate marker of nuclear translocation) was lower (p<0.01) in EPeri compared to other menopause stages; ERβ nuclear/cell protein ratio showed a similar trend (p=0.07). ER α/β protein ratio in whole cell showed no difference across stages, but was correlated with serum E2 (r=0.30, p=0.08) and estrone (r=0.38, p=0.03) across menopause stages. FMD was not correlated with any ER protein data.

CONCLUSION

The menopause transition, specifically the early perimenopausal period, may be a vulnerable period for the decline of endothelial ER α and β activation; however, we failed to show any correlation between endothelial function and ER protein content. Whether the decline in endothelial ER function/activity contributes to vascular dysfunction warrants further study.

Support or Funding Information

NIH R01AG049762, R01AG027678, R56HL114073, P30DK048520, P50HD073063, NCATS Colorado CTSA UL1TR001082, and Eastern Colorado VA GRECC

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Precision medicine in cow's milk allergy
Authors: D’Auria Enza , Venter Carina
Published: April 2020

Purpose of review

The aim of this review is to describe the role of precision medicine in the diagnosis, treatment, and monitoring of cow's milk allergy.

Recent findings

The development of ‘omics’ sciences in the field of food allergy has led to a better understanding of the allergenicity of cow's milk proteins and significant advances in the knowledge of the pathogenesis and mechanisms of cow's milk allergy. Omics-based technologies allow the practitioner to better differentiate cow's milk allergy subtypes and to predict cow's milk allergy (CMA) persistence over time. Precision medicine extends the role of the oral food challenge, to determine the individual's threshold doses, and to establish tolerance to baked milk products. Other than symptom relief, dietary strategies are currently being investigated for the potential to induce tolerance. Oral immunotherapy offers a treatment option for patients with severe and persistent IgE-mediated CMA. Individual baseline-immune profiles may be predictive of cow's milk oral immunotherapy safety and efficacy.

Patient data derived from current technology, in combination with the patient's history, can be translated into treatments targeted at patient-tailored interventions.

The identification of novel biomarkers may improve diagnostic accuracy and also predict patient responsiveness to treatments. Integration of patient data will become increasingly important as omics technologies become more widely used in the clinical setting.
Microbiota in non-IgE-mediated food allergy
Authors: Mennini Maurizio , Fierro Vincenzo , Di Nardo Giovanni , Pecora Valentina , Fiocchi Alessandro
Published: April 2020

Purpose of review

To perform a nonsystematic review of the literature on the microbiota in the different types of non-IgE-mediated food allergy.

Recent findings

The commonest non-IgE-mediated disorders managed by allergists include: eosinophilic esophagitis, food protein-induced enteropathy, food protein-induced enterocolitis syndrome, and food protein-induced allergic proctocolitis. The review of the literature describes how at phylum level we observe an increase of Proteobacteria in eosinophilic esophagitis esophageal microbiota and in food protein-induced enterocolitis syndrome, and food protein-induced allergic proctocolitis gut microbiota, while we observe an increase of Bacteroidetes in healthy controls. Several studies endorse the concept that a bloom of Proteobacteria in the gut reflects dysbiosis or an unstable gut microbial community structure. In several studies, the type of diet, the use of probiotics and in a single experience the use of fecal microbiota transplantation has produced significant variations of the microbiota.

Genetic factors alone cannot account for the rapid rise in food allergy prevalence and the microbiome might be contributing to allergy risk. Our review showed that common features of the pathological microbiota among different types of non-IgE-mediated food allergy can be identified. These evidences suggest a possible role of the microbiota in the pathogenesis and non-IgE-mediated food allergies and the need to understand the effects of its modulation on the disorders themselves.
Expert Consensus for a Principle-based Classification for Treatment of Diaphyseal Pediatric Femur Fractures
Authors: Weltsch Daniel , Baldwin Keith D. , Talwar Divya , Flynn John M.
Published: April 2020

Introduction:

Recent studies demonstrate considerable deviation from the American Academy of Orthopaedic Surgeons (AAOS) evidence-based guidelines for the treatment of pediatric diaphyseal femur fractures (PDFFs). This study aimed to determine if expert-consensus can be reached on a principle-based classification to be applied broadly to a wide variety of PDFF scenarios and if outcomes correspond to adherence to the classification.

Methods:

A 2-stage study was performed. First, a survey of experts using a principle-based approach to PDFF. We conducted a survey of 17 thought-leaders (criteria≥20 y’ experience+authors of the seminal pediatric femur fracture studies) who were asked to classify 15 cases of PDFF using the principle-based classification for agreement. Next, we conducted a retrospective review of 289 consecutive PDFF treated (2011-2015) at a level 1 pediatric trauma center. For each case, we compared the actual treatment and proposed “ideal” principle-based classification. We then compared clinical results and outcome data points including the length of stay, physician visits, and hospital charge data.

Results:

A substantial (κ=0.7) expert-agreement was noted for assigning treatment principles with near-perfect (κ=0.93) agreement on conservative versus surgical management. We obtained agreement on employing a flexible implant (κ=0.84) rigid fixation (κ=0.75) and damage control philosophy (κ=0.64). Suboptimal clinical results were noted in 43% of the undertreated patients (24/56), 18.8% of the adequately treated, and 14.3% of overtreated (P<0.01) patients. An increasing trend for the length of hospital stay and a number of clinic visits was noted as the treatment class increased (P<0.01). Charges were 4.2 times higher for an episode of operative versus nonoperative care (P<0.01). Rigid fixation (class 4) had significantly (P=0.01) higher total and material charges than flexible fixation (class 3).

Discussion:

The proposed classification has a substantial agreement among thought-leaders. Clinical results demonstrated significantly more suboptimal results in undertreated fractures, compared with ideally treated or more invasively treated fractures. More invasive treatments led to increased burden to families and the system in terms of length of stay and hospital charges.

Level of Evidence:

Level III.
Are Preoperative PHQ-9 Scores Predictive of Postoperative Outcomes Following Anterior Cervical Discectomy and Fusion?
Authors: Jenkins Nathaniel W. , Parrish James M. , Yoo Joon S. , Patel Dillon S. , Hrynewycz Nadia M. , Brundage Thomas S. , Singh Kern
Published: April 2020

Study Design:

This is a retrospective study.

Objective:

The objective of this study was to determine if there is an association between preoperative depression, as quantified by Patient Health Questionnaire-9 (PHQ-9), and postoperative improvement in pain and disability after anterior cervical discectomy and fusion (ACDF).

Summary of Background Data:

Few studies have quantified depression symptoms in the preoperative period using PHQ-9 and have tracked patient-reported outcomes (PROs) following ACDF.

Methods:

Patients undergoing ACDF were retrospectively reviewed and stratified by their preoperative PHQ-9 score. PROs, including Neck Disability Index (NDI), Visual Analogue Scale (VAS) neck and arm pain, and 12-Item Short Form (SF-12) Physical Component Score (PCS), were measured preoperatively and at 6-week, 3-month, 6-month, and 1-year postoperatively. PRO scores were analyzed amongst PHQ-9 cohorts using multiple linear regression. Achievement of minimum clinically important difference (MCID) was compared using χ2 analysis.

Results:

Higher PHQ-9 scores were associated with increased preoperative NDI, VAS neck, and VAS arm scores and significantly lower SF-12 PCS scores preoperatively. Cohorts experienced similar VAS pain scores up to 1-year following surgery, except for VAS neck pain at 3 months when patients with greater depression symptoms had more pain. High PHQ-9 patients had higher NDI values at 6 weeks and 3-month marks but had similar NDI scores at 6 months and 1-year. Similarly, SF-12 PCS scores were lower for patients with a higher PHQ-9 score at 3 and 6 months, however, both groups had similar scores at 1-year follow-up. A greater percentage of the high PHQ-9 cohort achieved MCID for NDI, however, there were no differences in MCID achievement for VAS neck, VAS arm, or SF-12 PCS.

Conclusions:

Patients with worse preoperative mental health reported significantly greater preoperative disability and pain. However, both cohorts demonstrated similar clinical recovery at the 1-year follow-up. These findings suggest patients with worse preoperative mental health can expect significant improvements in PROs following surgery.
Comparison of IV Insulin Dosing Strategies for Hyperkalemia in the Emergency Department
Authors: Moussavi Kayvan , Nguyen Lani T. , Hua Henry , Fitter Scott
Published: April 2020

Objectives:

The objectives of this study were to evaluate the safety and efficacy of insulin dosing of less than 10 units versus 10 units in patients receiving hyperkalemia treatment.

Design:

Retrospective single-center study.

Setting:

Emergency department at a large academic medical center in the United States.

Patients:

Seven hundred adults treated for hyperkalemia with IV regular insulin between April 1, 2013, and September 27, 2018.

Interventions:

Patients that received less than 10 units of insulin were compared to those that received 10 units of insulin.

Measurements and Main Results:

Patients treated with less than 10 units had significantly lower frequency of hypoglycemia (11.2% vs 17.6%; p = 0.008). Reduction in serum potassium was significantly more modest in size in patients treated with less than 10 units (mean reduction 0.94 ± 0.71 mMol/L) compared with patients treated with 10 units (mean reduction 1.11 ± 0.8 mMol/L; p = 0.008). There were no statistically significant differences between groups in time to hypoglycemia, nadir serum glucose, severe hypoglycemia (<40 mg/dL), dextrose requirements, use of concurrent agents for hyperkalemia, need for repeat insulin dosing, length of stay, or mortality.

Conclusions:

Patients treated for hyperkalemia with insulin doses less than 10 units had reduced frequency of hypoglycemia; however, potassium reduction post treatment was more modest in these patients. These findings suggest providers choosing to administer 10 units IV insulin should ensure patients have adequate monitoring for hypoglycemia.
Effect of Very Low-Dose Hydrocortisone on Shock Reversal in Patients With Septic Shock
Authors: Sbertoli Robert , Hu Zeyu , Henke Jonathan , Wu Eric , Santosh Shrihari , Osmon Stephen , Charbek Edward , Jamkhana Zafar , Santosh Sadashiv
Published: April 2020

Objectives:

In patients with septic shock, hydrocortisone 200–400 mg/d has been shown to reverse shock compared with placebo. Lower doses of hydrocortisone have not previously been studied, and there are no previous studies comparing two different doses of hydrocortisone. At our institution, some clinicians routinely prescribe doses less than 200 mg/d. This study aims to compare the effect of lower doses of hydrocortisone to standard doses on shock reversal and adverse events in septic shock.

Design:

Retrospective cohort study.

Setting:

Single-center medical ICU.

Subjects:

Patients who received hydrocortisone for septic shock.

Interventions:

Electronic chart review.

Measurements and Main Results:

Patients were divided into low-dose hydrocortisone (75–150 mg/d) and standard-dose hydrocortisone (200–400 mg/d) cohorts based on initial prescribed hydrocortisone dose. Rates of shock reversal and adverse events in the two cohorts were compared. Two-hundred thirteen patients were included—41 in low-dose and 172 in standard-dose cohorts. Baseline characteristics including initial vasopressor requirement and Sequential Organ Failure Assessment scores were similar. Average rates of change in vasopressor needs, conditional hazard rate for vasopressor withdrawal, and cumulative probability for vasopressor withdrawal were all quantitatively similar for low-dose and standard-dose hydrocortisone. Insulin requirement (particularly in those with diabetes mellitus), blood glucose in those with diabetes mellitus, and frequency of secondary infections seemed to be lower in the low-dose hydrocortisone cohort. Mortality and other secondary outcomes were similar.

Conclusions:

In septic shock, hydrocortisone dosed 75–150 mg/d appears to reverse shock as effectively 200–400 mg/d and may cause a lower frequency of adverse events.
Long non-coding RNA small nucleolar RNA host gene 6 aggravates pancreatic cancer through upregulation of far upstream element binding protein 1 by sponging microRNA-26a-5p
Authors: Zhang Xing-Xing , Chen Hua , Li Hui-Ying , Chen Rui , He Lei , Yang Juan-Li , Xiao Lin-Lin , Chen Jin-Lian
Published: April 2020

Background

Pancreatic cancer (PC) is a highly deadly malignancy with few effective therapies. We aimed to unmask the role that long non-coding RNA small nucleolar RNA host gene 6 (SNHG6) plays in PC cells by targeting far upstream element binding protein 1 (FUBP1) via microRNA-26a-5p (miR-26a-5p).

Methods

SNHG6expression was predicted by bioinformatics, followed by verification via reverse transcription quantitative polymerase chain reaction. Then, the interactions amongSNHG6,miR-26a-5p, andFUBP1were detected through online software analysis, dual luciferase reporter assay and RNA pull-down. After that, cells were treated with different small interfering RNAs and/or mimic to determine the interactions amongSNHG6,miR-26a-5p, andFUBP1and their roles in PC cells. Finally, the role ofSNHG6in tumor growthin vivowas evaluated by measuring the growth and weight of transplanted tumors in nude mice. At-test, one-way and two-way analysis of variance were used for data analysis.

Results

Compared with that in normal tissues,SNHG6was highly expressed in PC tissues (1.00 ± 0.05vs.1.56 ± 0.06,t = 16.03,P < 0.001). Compared with that in human pancreatic duct epithelial cells (HPDE6-C7),SNHG6showed the highest expression in PANC-1 cells (1.00 ± 0.06vs.3.87 ± 0.13,t = 34.72,P < 0.001) and the lowest expression in human pancreatic cancer cells (MIAPaCa-2) (1.00 ± 0.06vs.1.41 ± 0.07,t = 7.70,P = 0.0015). Compared with the levels in the si-negative control group,SNHG6(0.97 ± 0.05vs.0.21 ± 0.06,t = 16.85,P < 0.001), N-cadherin (0.74 ± 0.05vs. 0.41 ± 0.04,t = 8.93,P < 0.001), Vimentin (0.55 ± 0.04vs.0.25 ± 0.03,t = 10.39,P < 0.001), and β-catenin (0.62 ± 0.05vs.0.32 ± 0.03,t = 8.91,P < 0.001) were decreased, while E-cadherin (0.65 ± 0.06vs.1.36 ± 0.07,t = 13.34,P < 0.001) was increased afterSNHG6knockdown ormiR-26a-5poverexpression, accompanied by inhibited cell proliferation, migration, and invasion.SNHG6overexpression exerted the opposite effects.SNHG6upregulatedFUBP1expression by spongingmiR-26a-5p. SilencingSNHG6blocked the growth of PCin vivo.

Conclusion

SilencingSNHG6might ameliorate PC through inhibition ofFUBP1by spongingmiR-26a-5p, thus providing further supporting evidence for its use in PC treatment.
Evaluation of an Educational Intervention Utilizing Nurse “Champions” and Nurses' Documentation of Intensive Care Unit Delirium
Authors: DeForge Christine E. , Yip Natalie H. , Dzierba Amy L. , Ryan Patrick G. , Larson Elaine L.
Published: April 2020

Background

Delirium is a common, underdetected problem that has short- and long-term negative sequelae for critically ill patients. Prompt and accurate delirium identification by nurses can ensure early intervention and treatment to help minimize adverse outcomes.

Objectives

To evaluate the relationship between an educational program and the accuracy of registered nurses' (RNs') documentation of the Confusion Assessment Method for the Intensive Care Unit (CAM-ICU), a delirium screening tool.

Methods

In a medical ICU at a tertiary academic medical center from September 2015 to March 2016, RNs were reinstructed on use of the CAM-ICU. Registered nurse assessment data were collected retrospectively for 12 months before and after intervention and were compared against the CAM-ICU algorithm using χ2 analysis.

Results

A total of 10 736 RN assessments in 1020 patients preintervention and 11 068 in 951 patients postintervention were evaluated. Overall RN accuracy improved from 78% to 80% (P = .054). The algorithm determined delirium to be present in 32% versus 30% of all patients preintervention and postintervention, respectively; there was no difference in rate of nurse detection of delirium preintervention and postintervention (54% vs 55%, not statistically significant). The percentage of “inappropriate unable to assess” ratings by nurses decreased from 42% to 37% postintervention (P < .05).

Conclusions

After a comprehensive training initiative, there was no significant improvement in CAM-ICU documentation and no improvements in patient delirium identification. Future quality improvement efforts should target reducing the number of assessments that RNs judge to be “unable to assess.” Clinical practice must evolve to routinely incorporate RN delirium assessments into the patient's plan of care.