bluuebunny/crossref_metadata_2025_split huggingface dataset:
Depletion of Caveolin‐1 in Lung Vasculature and Increase in Caveolin‐1 Positive Circulating Extracellular Vesicles as Early Biomarkers of Endothelial Injury
Authors: Oliveira Suellen Darc Santos , Castellon Maricela , Machado Roberto F , Elliott Michael H , Bonini Marcelo G , Minshall Richard D 
Published: June 2021

RationaleEndothelial cell activation and dysfunction are hallmark features of Acute Lung Injury and Acute Respiratory Distress Syndrome (ALI/ARDS). Caveolin‐1 (Cav‐1) is highly expressed in pulmonary microvascular endothelial cells (PMVECs) and plays a key role in maintaining vascular homeostasis. The aim of this study was to determine whether the lung inflammatory response to endotoxin promotes PMVEC Cav‐1 depletion and appearance of Cav‐1 positive circulating extracellular vesicles (Cav‐1+ EVs) and whether this contributes to ALI.MethodsALI was induced in C57BL6 WT mice (male; 2.5–3 months old) by nebulized Escherichia coli lipopolysaccharide (LPS; 10 mg over 1 hr daily for 1 or 4 days). Lung pathology, Cav‐1 and eNOS expression, Cav‐1 ubiquitination, and Cav‐1+ EVs were quantified in total lung or platelet‐free plasma by immunoshistochemistry and western blot, respectively. Broncho‐alveolar lavage fluid (BALF) obtained from control and LPS‐treated mice was used to measure inflammatory cell, cytokine, and albumin levels.ResultsAlveolar accumulation of plasma albumin, immune cells, and pro‐inflammatory and regulatory cytokines (TNF‐α, IL‐1β, IL‐6, TGF‐β) was signficantly elevated in LPS‐treated WT mice. Intermittent airway LPS exposure for 96 hrs also increased Cav‐1 phosphorylation and ubiquitination (Ub) and decreased total lung Cav‐1 expression by 48 ± 4.16 % (n=3–4 mice; P<0.05) when compared to untreated WT mice. Depletion of EC Cav‐1 was accompanied by a decrease in eNOS dimerization indicative of endothelial dysfunction. In addition, circulating Cav‐1+ EVs increased by 38% after 24 hrs of LPS exposure (0.61 ± 0.05 to 0.98 ± 0.16 a.u.) in control and LPS‐treated mice, respectively (n= 8–10 mice; P<0.05) and decreased below basal level after 96 hrs indicating PMVEC Cav‐1 depletion may be due to both Cav‐1 ubiquitination and proteosomal degradation as well as release Cav‐1+ EVs into the circulation. PMVEC Cav‐1 depletion in response to endotoxin exposure was associated with endothelial dysfunction and vascular injury suggesting EC Cav‐1 depletion may play a significant role in the onset of ALI. Similar lung pathology was observed in EC‐specific Cav‐1−/− mice.ConclusionEndothelial cell Cav‐1 depletion and Cav‐1+ EVs may be an early biomarker of pulmonary vascular injury that may predict pulmonary vascular inflammatory disease due to endothelial dysfunction.Support or Funding InformationFinancial support: CNPq Fellowship‐CsF/Brazil (SDO) and HL125356 and DOD W911NF‐15‐R‐002 (RDM, MGB).
Prolonged Heat Stress Altered Autophagy Signaling in Oxidative Skeletal Muscle
Authors: Selsby Joshua , Ganesan Shanthi , Brownstein Alexandra J , Gabler Nicolas K , Pearce Sarah C , Baumgard Lance H , Rhoads Robert P 
Published: June 2021

Prolonged heat stress represents a continuing threat to human health and agricultural production economics. Despite this broad, negative impact little is known about underlying pathological mechanisms, which has limited the development of etiological interventions and left clinicians and producers with only cooling and rehydration strategies. The purpose of this investigation was to determine the extent to which prolonged heat stress altered autophagic signaling in oxidative skeletal muscle. We hypothesized that heat stress would induce autophagy independent of the volitional caloric restriction that accompanies heat stress. To test this hypothesis pigs were treated as follows: thermoneutral (TN; 20°C), heat stress (HS; 35°C), or were house in thermoneutral conditions but pair‐fed to the heat stress group (PFTN) for seven days. Upon euthanasia the red portion of the semitendinosus muscle was collected for subsequent measures of relative protein abundance. We determined that markers of autophagy initiation, including phosphorylated adenosine monophosphate activated protein kinase (AMPK) and Unc51‐like kinase 1 (Ulk1), were increased approximately 3‐fold (p<0.05) in HS compared to TN and PFTN. Similarly, nucleation markers, PI3 kinase class III and Beclin‐1, were increased approximately 3‐fold (p<0.05) in HS compared to TN and PFTN. We also discovered that elongation markers autophagy related gene (Atg)12 and Atg16 were increased 2‐ and 4‐fold (p<0.05), respectively, in HS compared to TN and PFTN, while Atg5 was similar between groups. Markers of autophagosome formation including microtubule‐associated protein 1A/1B‐light chain (LC3) II and the LC3 II/I ratio were increased 5‐fold (p<05) in HS and PFTN compared to TN while LC3 I was similar between groups. Sequestosome 1 (SQSTM1/p62) is consumed during autophagosomal degradation by the lysosome and is thus an inverse correlate of autophagic degradation. We discovered that relative protein abundance of p62 was increased 5‐fold (p<0.05) in HS compared to TN and PFTN. In partial support of our hypothesis, these data indicate heat stress‐mediated induction of autophagic signaling is largely independent of reduced feed intake. Further, these data also suggest a HS‐mediated stalling of autophagic degradation despite prolonged reduced feed intake.Support or Funding InformationThis work supported by USDA grants 2014‐67015‐21627 and 2011‐67003‐30007.
Measurement of the Compensatory Reserve Predicts Racial Differences in Orthostatic Tolerance
Authors: Wenner Megan M. , Hinds Kumba , Howard Jeffrey T , Nawn Corrine D , Stachenfeld Nina S , Convertino Victor A 
Published: June 2021

The Compensatory Reserve Index (CRI) has recently been established to accurately measure the body's integrated capacity to compensate for physiological conditions of reduced central blood volume, and predict hemodynamic decompensation. We previously demonstrated that African American (AA) women have a higher tolerance to orthostatic stress. Therefore, we tested the hypothesis that the CRI would identify racial differences in orthostatic tolerance prior to the onset of traditional signs and symptoms. We measured beat‐by‐beat blood pressure (BP) and heart rate (HR) in 23 AA (22 ± 1 years; 24 ± 1 kg/m2) and 31 Caucasian women (20 ± 1 years; 23 ± 1 kg/m2) during progressive lower body negative pressure (LBNP) to presyncope. Blood pressure waveforms were analyzed using a machine‐learning algorithm to derive the CRI at each LBNP stage; stroke volume (SV) was derived from the Finometer (Model flow). Resting mean arterial BP (AA, 78 ± 3 vs. Caucasian, 74 ± 2 mmHg) and HR (AA, 68 ± 2 vs. Caucasian, 65 ± 2 bpm) were similar between groups. The CRI progressively decreased during LBNP in both groups, however the CRI in Caucasian women was 4% lower than AA women at −15 mmHg LBNP and decreased to 21% lower at −50 mmHg LBNP (P<0.05). The rate of decline in SV during LBNP was greater in Caucasian women (ANOVA P<0.05), and differences were noted as early as −20 mmHg. However changes in BP and HR during LBNP were not different between groups. At presyncope, mean arterial BP (AA, 66 ± 3 vs. white, 65 ± 4 mmHg) and HR (AA, 102 ± 4 vs. white, 97 ± 3 bpm) were similar between groups, as was the change in SV (AA, Δ −33 ± 2 vs. white, Δ −35 ± 3 mL). Finally, the time to presyncope was 1.6 minutes longer in AA women. These data support the notion that greater tolerance to the orthostatic stress induced by LBNP in AA women can be explained by their greater reserve to compensate for progressive central hypovolemia compared to Caucasian women. An important clinical translation is that the CRI provides a more sensitive and specific measure for early assessment of differences in orthostatic tolerance in comparison to traditional vital signs and symptoms or changes in hemodynamics.Support or Funding InformationResearch supported by funding from the Combat Casualty Care Research Program of the US Army Medical Research and Materiel Command and NIH HL071159.
Determination of the Bioactive Components in a Grape Seed Procyanidin Extract Responsible for Enhanced Farnesoid X Receptor Transactivation
Authors: Rodriguez Kelvin , Ricketts Marie‐Louise 
Published: June 2021

Dietary procyanidins are beneficial for maintaining health. Grape seed procyanidin extract (GSPE) in particular has been shown to regulate cholesterol and triglyceride (TG) homeostasis in vivo (1–5). We have identified several underlying molecular mechanisms by which GSPE decreases serum TG levels, including farnesoid x receptor (FXR)‐dependent decreased hepatic triglyceride synthesis, inhibition of enterohepatic bile acid recirculation and increased hepatic TG catabolism (3–5). Furthermore, we identified GSPE as a histone deacetylase (HDAC) inhibitor, leading to enhanced peroxisome proliferator‐activated receptor alpha (PPARα) target‐gene transcription and increased fatty acid β‐oxidation (6). During our initial investigations we discovered that GSPE acts as a co‐agonist ligand for FXR (2). The present study was designed as a follow‐up to this initial observation and aimed to identify which component (or components) within GSPE are responsible for the observed FXR‐dependent co‐agonist activity. GSPE was fractionated by column chromatography to separate compounds based on molecular size (n=3). The degree of polymerization for each fraction was determined by high performance liquid chromatography. Using a cell‐based transient transfection assay, each fraction was tested in combination with the bile acid, chenodeoxycholic acid (CDCA), to assess its ability to increase FXR transactivation (n=3, analyzed in triplicate), and compared to the effect seen with GSPE. Five different fractions from each column consistently enhanced FXR transactivation and were subsequently analyzed by mass spectrometry (MS). The MS results show that the fractions that enhance FXR transactivation are enriched in dimers and dimer gallates. Ultimately, this study facilitates enhanced understanding regarding the observed GSPE‐mediated FXR‐dependent regulatory effects on metabolic homeostasis observed in vivo.Support or Funding InformationUSDA NIFA Hatch Project NEV0738; USDA NIFA Multi‐state Project W‐3122: Beneficial and adverse effects of natural chemicals on human health and food safety; USDA NIFA Hatch Project NEV0749; A grant from the National Institute of General Medical Sciences (P20GM103440) from the National Institutes of Health.
Knockout of the PHLDA1 gene in breast cancer cells reveals multiple roles for PHLDA1 in cancer phenotypes
Authors: Zimnicka Adriana M. , Sharma Tiffany , Regan Maureen , Merrill Brad J. , Frasor Jonna 
Published: June 2021

PHLDA1 or pleckstrin‐homology‐like domain family A, member 1, also known as TDAG51, is a small, PH‐domain containing protein that acts as a potential modulator of apoptosis and cell proliferation in breast carcinomas. However, many contradictory findings have been reported and the mechanisms by which PHLDA1 affects cancer cells are not well defined. We have reported that in estrogen receptor (ER)‐positive breast tumors, high PHLDA1 mRNA expression was found to be associated with a high risk of distant metastasis, while in ER negative breast tumors, high PHLDA1 expression coincided with a more favorable outcome, suggesting that PHLDA1 has opposite effects depending on the ER status of a tumor. We have recently shown that PHLDA1 contributes to the survival and growth of breast cancer stem cells in ER positive mammospheres, where PHLDA1 mRNA and protein are regulated directly by a crosstalk between ER and the inflammatory nuclear factor κB (NFκB) pathway, and indirectly through inhibition of miR‐181a and b, which targets PHLDA1 for degradation. Here, using CRISPR/Cas9 mediated genome editing, we have permanently knocked out PHLDA1 gene from ER positive MCF‐7 and ER negative MDA‐MB‐231 breast cancer cell lines, and characterized their phenotype. While MDA‐MB‐231 cells are still under investigation, here we report that MCF‐7 cells missing the PHLDA1 gene showed decreased cell proliferation, migration, and invasion across a matrigel layer, consistent with a role for PHLDA1 in more aggressive ER+ breast cancers. In contrast, PHLDA1‐knock out cells were less susceptible to apoptosis than control cells, as shown by decreased activity of caspase‐3 and −7, and were rendered more resistant to doxorubicin and paclitaxel‐induced cytotoxicity, suggestive of a pro‐apoptotic role for PHLDA1. This phenotype was reversed by overexpressing PHLDA1 in the PHLDA1 (−/−) cells. We also compared PI3K activation and Akt phosphorylation in parental and knock out cells in order to determine the signaling pathway mediating the effects of PHLDA1. The PH domains, or phosphoinositide binding protein modules, are known to govern the function of two other PHLDA isoforms, PHLDA2 and PHLDA3, but no information on the role of PH domain in the function of PHLDA1 has been described. We created a PH domain deletion mutant (PHLDA1‐ΔPH), and using confocal microscopy assessed its subcellular localization, as well as Akt/PI3K signaling pathway activity, and compared it to the WT PHLDA1. Our study sheds new light on the function of PHLDA1 in breast cancer cells, suggesting a broad role as an anti‐proliferative/anti‐invasive but pro‐apoptotic protein. Moreover, we describe the signaling pathway involving PHLDA1, an understanding of which will potentially define novel therapeutic targets in treatment of breast carcinomas.Support or Funding InformationThis study was supported by the NIH fund 1 R01 CA200669 01 and DOD fund W81XWH‐16‐1‐0044 to JF
Insight into the Evolution of the Avian Vocal Organ, or Syrinx, from Enhanced‐Contrast X‐ray Computed Tomography and Fossil Data
Authors: Clarke Julia , Li Zhiheng , Riede Tobias , Goller Franz 
Published: June 2021

From complex songs to simple honks, squawks, or caws, birds produce sounds using a unique vocal organ, the syrinx. Located close to the heart at the tracheobronchial junction, vocal folds or membranes attached to modified mineralized rings vibrate to produce sound. Not thought to commonly enter the fossil record, the few reported fossilized parts of the syrinx are geologically young (i.e., Pleistocene and Holocene). The only known older syrinx is an Eocene specimen, which was not described or figured. Data on the relationship between soft tissue structures and syringeal three‐dimensional geometry has also been limited. Our team of physiologists and paleontologists recently described the first remains of a fossil syrinx from the Mesozoic, which are preserved in three dimensions in a specimen from the Late Cretaceous of Antarctica. With both cranial and postcranial remains, the new specimen is the most complete to be recovered from a part of the radiation of living birds, Aves. Enhanced‐contrast x‐ray computed tomography (CT) of syrinx structure in twelve extant non‐passerine birds, as well as CT imaging of the Cretaceous and the Eocene syrinxes, informs both the reconstruction of ancestral states in birds and properties of the vocal organ in the extinct species. Fused rings in the Cretaceous fossil form a well‐mineralized pessulus, a derived neognath bird feature, proposed to anchor enlarged vocal folds, or labia. Left‐right bronchial asymmetry, only known in extant birds with two sound sources is similarly present.The new data show the fossilization potential of the avian vocal organ and beg the question why these remains have not been found in other dinosaurs. The lack of other Mesozoic tracheal and bronchial ring remains, and the poorly‐mineralized condition in archosaurian taxa without a syrinx, may indicate a complex syrinx was a late arising feature in the evolution of birds, well after the origin of flight and respiratory innovations. While a role for complexity in social structure and mating system has been extensively investigated with reference to increases in brain size in other vertebrates including humans, in the early evolution of birds these hypotheses have not been explored. We propose that first more complex visual signals and then vocal communication and song may have played important roles in the evolution of the larger extant avian brain.Support or Funding InformationGordon and Betty Moore Foundation Grant GBMF4498, US National Science Foundation (OPP ANT‐1141820, OPP 0927341 and EAR 1355292 to J.C.); Argentine Agencia Nacional de Promoción Científica y Técnica (PICT 2010‐066; F.E.N).
Behavior related genes, dietary preferences and anthropometric traits
Authors: Berciano Silvia , Lai Chao‐Qiang , Herranz Jesus , Aslibekyan Stella , Claas Steve A , Irvin Marguerite R , Tsai Michael Y , Hopkins Paul N , Hidalgo Bertha , Arnett Donna K , Ordovas Jose M , Tiwari Hemant 
Published: June 2021

Food preferences and cognitive control influence dietary habits thus affecting compliance with dietary advice and the risk of chronic diseases. A complex mix of genetic and environmental, cultural and social factors drives these preferences. Therefore, efforts to improve diet and behavior at the individual level should take into consideration this distinct combination of genetic and environmental factors, specifically addressing the psychological component of food consumption with the goal of facilitating long‐term compliance with dietary interventions.ObjectiveTo investigate the association between behavioral candidate genes, food preferences and anthropometric traits.Population and MethodsFor this purpose we used data from the Genetics and Lipid Lowering Drugs and Diet Network (GOLDN) Study including two genetically homogeneous sites (Minneapolis, Minnesota, and Salt Lake City, Utah). In the current study, 818 participants (404 men and 414 women) of European ancestry were included in our analyses. Single nucleotide polymorphisms (SNPs) within 38 loci (1359 SNPs) selected on the basis of previous associations with several behavioral and psychological traits (i.e., stress, addiction, depression, impulsivity, novelty‐seeking, aberrant eating) were extracted from the original genome genotype data that was generated from the Genome‐Wide Human SNP Array 6.0 (Affymetrix). Information about dietary intake was collected with the use of a diet‐history questionnaire, which was developed by the National Cancer Institute.ResultsMultiple nominally significant associations (p<0.05) were observed between genetic variability at the selected loci and the consumption of specific foods and nutrients. However, after adjustment for multiple comparisons, significant associations (Padj<0.05) were observed for the FTO locus with vegetable and total fiber intake; the CREB1 and GABRA2 loci were associated with salt intake; and the SLC6A2 with total fat and monounsaturated fatty acids. Finally, chocolate intake was associated with variation at the OXTR locus. The most significant association with anthropometric traits was found for OXTR and waist circumference.ConclusionOur data indicate that genes implicated in behavioral and psychological traits drive a significant component of an individual's food preferences and dietary habits. This information will contribute to a better understanding of eating behavior and facilitate the implementation of personalized dietary advice that should result in better compliance and more successful prevention and therapy of chronic disorders.Support or Funding InformationNational Heart, Lung, and Blood Institute (NHLBI) grants U01HL072524 (Genetic and Environmental Determinants of Triglycerides), NHLBI R01 HL091357 and by contracts 53‐K06‐5‐10 and 58‐1950‐9‐001 from the USDA, Agriculture Research Service
A Genome‐Wide Analysis of Placental Epigenetic Markers in Response to Maternal Choline Supplementation
Authors: Kwan Sze Ting (Cecilia) , Wang Minghui , King Julia H , Yan Jian , Jiang Xinyin , Sun Qi , Roberson Mark S , Caudill Marie A 
Published: June 2021

Choline is an essential micronutrient that provides methyl groups for cellular DNA methylation, an epigenetic modification with downstream effects on gene expression. Maternal choline supplementation (MCS) during pregnancy has many beneficial effects on placental and fetal development, which may be mediated in part by an altered placental methylation profile. In addition to DNA methylation, other epigenetic mechanisms such as genomic imprinting and microRNA (miRNA) regulation are also important to normal placental and fetal development, but whether these markers are modulated by choline remains largely unknown. We sought to investigate the impact of MCS on these epigenetic markers using a subset of placental samples collected at gestational day 15.5 from dams consuming 1× or 4× the recommended choline level. Placental mRNA and miRNA were extracted and sequenced, and the RNAs with differential expression were determined using the edgeR package developed in R. In response to MCS, three imprinted genes (Dcn, Qpct and Tnfrsf23) were downregulated in the male placentas (Padj≤0.01) whereas four imprinted genes (Ampd3, Tfpi2, Gatm, and Aqp1) were upregulated in the female placentas (Padj≤0.05). Furthermore, five miRNAs (miR‐712‐5p, miR‐3470a, miR‐6538, miR‐6240 and miR‐5126) were significantly downregulated in the female placentas from the 4× choline group (Padj≤0.033). Consistent with the downregulation of these miRNAs, some of their predicted mRNA targets showed a higher expression (P<0.05). These target genes are involved in processes such as cell signaling, immune response and macronutrient transport, all of which are crucial to placental development and fetal health. In contrast, none of the miRNAs in the male placentas remained significant after correction for false discovery rate (Padj>0.05). Overall, these data indicate that the placental imprintome and miRNA profile are responsive to MCS, but the effects may differ between the male and the female placentas.Support or Funding InformationUSDA, NIH, Graduate Women in Science, Egg Nutrition Center