bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transcriptomic Analysis Reveals Different Sensitivities of Genes to EP4 Receptor Agonist Stimulation in BEAS‐2B Human Bronchial Epithelial Cells
Authors: Joshi Radhika , Yan Dong , Hamed Omar , Giembycz Mark A. 
Published: April 2020

IntroductionProstanoid EP4‐receptors are highly expressed on human airway epithelial cells (AECs) and typically mediate canonical, Gs‐cAMP signaling. EP4‐receptor agonists exhibit bronchodilator and anti‐inflammatory activity and could represent novel therapeutic candidates to treat obstructive lung diseases. We have reported previously that long‐acting β2‐adrenoceptor agonists (LABAs) promote significant gene expression changes in human AECs, which could contribute to their clinical efficacy. In this study, we have determined the EP4‐receptor‐regulated transcriptome in BEAS‐2B cells using the selective agonist, ONO‐AE1‐329, using the LABA, vilanterol, as a comparator.MethodsBEAS‐2B cells were treated for 2h with vehicle, and two concentrations of ONO‐AE1‐329 (1nM, ONO1; 1000nM, ONO1000). A maximally effective concentration of vilanterol (100nM, Vil100) was included to define maximum responses. Gene expression changes were determined by RNA sequencing. Simulated E/[A] curves were generated from ONO1 and ONO1000 data assuming a unity Hill coefficient for each gene expression change (≥3‐fold; FDR≤5%). BEAS‐2B cells transfected with a cAMP response element (CRE) luciferase reporter were used to determine the affinity (KA) of ONO‐AE1‐329 for the EP4‐receptor. This was estimated by operational model fitting of E/[A] curve data before and after fractional receptor depletion using siRNA mediated PTGER4 gene silencing. Occupancy‐response relationships for ONO‐AE1‐329‐induced genes were constructed using the simulated E/[A] curves and the KA value determined in CRE reporter cells.ResultsTranscriptomic changes produced by Vil100 and ONO1000 were highly correlated suggesting that the β2‐adrenoceptor and the EP4‐receptor shared a common mechanism of action. Vilanterol was a full agonist on all induced genes whereas ONO‐AE1‐329 was a partial agonist with intrinsic activity values that varied between genes. The KA of ONO‐AE1‐329 was found to be consistent with a previous radioligand binding study. ONO‐AE1‐329‐induced genes differed in sensitivity by ~18‐fold with NR4A3 and PDE4B representing transcripts at the extremes of this sensitivity spectrum. Accordingly, receptor occupancy‐response relationships varied from almost linear (where response is directly proportional to occupancy; e.g., NR4A3) to markedly hyperbolic (indicative of a significant receptor reserve; e.g., PDE4B) (Fig.
1).ConclusionsDifferences in the sensitivities of genes to ONO‐AE1‐329 indicate that the EP4‐receptor interprets equivalent degrees of receptor occupancy differently. This may be explained by ‐ a) regulation by multiple cAMP‐induced transcription factors, and/or b) variability in promoter context and co‐factor binding which may differentially affect the ability of transcription factors to interact favorably with DNA.Support or Funding InformationCIHR‐ PJT 152904Receptor occupancyresponse relationships for ONO‐AE1‐329 induced genesFigure 1
Zanthoxylum piperitum activates thermogenic gene expression and beige adipogenesis in white adipose tissues of mice
Authors: Ihara Hayato , Takeshima Ken , Kadowaki Akio , Gato Nobuki , Kishida Kunihiro , Akamizu Takashi 
Published: April 2020

Metabolic syndrome increases the risk of cardiovascular disease, stroke, and diabetes mellitus. Because treatment of obesity is necessary to prevent multiple organ dysfunctions associated with metabolic syndrome, tolerable anti‐obesity drugs must be developed. In this study, we focused on Zanthoxylum piperitum (ZP), a uniquely flavored spice that confers a tingling sensation. We examined the effects of ZP on obesity and glucose–lipid metabolism in mice fed a standard diet (STD) or high‐fat diet (HFD) with or without ZP. Notably, body weight and fat mass markedly decreased, and glucose–lipid metabolism improved, in STD‐fed mice treated with ZP. Moreover, thermogenic gene expression was elevated in inguinal white adipose tissue (iWAT) from STD‐fed mice treated with 3% ZP. Immunohistochemistry of iWAT revealed uncoupling protein‐1 (UCP‐1)–positive multilocular adipocytes, suggesting induction of the beige adipogenesis. In conclusion, ZP administered in the diet decreases weight and fat mass, concomitant with induction of beige adipogenesis. These findings imply that identification of the components of ZP that exert anti‐obesity effects could facilitate development of novel anti‐obesity drugs originating from a natural food.Support or Funding InformationThis work was supported by JSPS KAKENHI Grant Number JP25504010 (to HI), JP16K12728 (to HI), Grants‐in‐Aid from the Consortium of Higher Education Institution in Wakayama (2012&2016 to HI, 2014&2015 to KT), Research and Development Support Project for Pioneering Industrial Technology by Wakayama Prefectural Government (to NG), and a Grant‐in‐Aid from the Regional Innovation Strategy Support Program 2011–2016 (to KT).Visceral and subcutaneous fat mass were quantified by small‐animal CTFigure 1H&E staining and immunohistochemistry showed UCP‐1 positive multilocular adipocytes in iWAT from STD+3%ZP miceFigure 2
Locating TATA Binding Protein in the RNA Polymerase I Pre‐Initiation Complex
Authors: Vo Giang , Jackobel Ashleigh , Knutson Bruce 
Published: April 2020

Transcription of eukaryotic DNA into RNA is carried by three main RNA polymerases (Pols I–III). Pol I synthesizes ribosomal DNA (rDNA) into ribosomal RNA (rRNA), which has been directly linked to the rate of protein synthesis, cell growth and proliferation. The efficiency of Pols’ function is controlled by general transcription factors, which enable the enzyme to interact with promoter DNA at the initiation stage. In Saccharomyces cerevisiae, Pol I transcription initiation relies on four main factors, one of which is TATA Binding Protein (TBP). TBP is an essential universal transcription factor used by all three eukaryotic Pols. As its name eludes, TBP typically binds to TATA motifs found in promoter DNA. Interestingly, unlike Pol II and III, Pol I promoters are TATA‐less. Thus, unlike the location of TBPs in other PICs, the location and function of TBP within the Pol I PIC remains unclear.Current Cryo‐EM structures of CF‐DNA‐Pol I initiation complex leaves little room for TBP to bind to its “typical” Pols II/III position. Therefore, we hypothesize that TBP must occupy a unique location, putatively more upstream, within the Pol I PIC. Combining available structural data with previous crosslinking data of CF and TBP, we modeled the interaction between the two proteins with varying distance constraints. In addition, we analyzed TBPs location on the rDNA promoter by site‐directed hydroxyl radical cleavage experiment, known as FeBABE, where DNA phosphothioate modification probes were constructed based on our predicted models. Our results from these combinational methods elucidated that TBP locates more upstream within Pol I PIC.Support or Funding InformationThis work was a part of SUNY Upstate Medical School’s Summer Undergraduate Research Fellowship 2019 and was supported by National Cancer Institute (NCI), National Institute for Dental and Craniofacial Research (NIDCR), and Carol Baldwin Research Foundation.FeBABE site‐directed hydroxyl radical cleavage.A and B. Sites of FeBABE incorporation into rDNA promoter in clusters of 5 include the “typical” location of TBP (1), upstream of the CE (2), 5′ of CE (3), and within the CE (4). C. Western blot showed FeBABE specific cleavage products. Cleavages were observed when FeBABE was incorporated at all four positions. Most intense cleavages were observed when FeBABE is incorporated upstream of the CE. D. Quantitation of TBP cleavages.Figure 1
Disorders of the metabolic status and morphofunctional state of liver and kidneys of chicken
Authors: Anipchenko Polina , Shabunin Sergey , Kotarev Viacheslav , Mikhailov Evgenii , Stekol'nikov Anatolii 
Published: April 2020

In this study we analyzed biochemical parameters of blood, tibia, histological analysis of liver and kidney tissues of 46‐days old pullets of egg‐laying chickens. The research aim is study of biochemical and morphological changes in replacement pullets to predict risks of growth weak replacement pullets. The research shows that in the serum total proteins were higher then normal range on 28, 8%, albumins on 32,0%. High level of this parameters leads to produce too much uric acid and accumulate it in the body and its higher on 21,3%. Energy change in serum is shown with satisfying range of glucose and reduced rate of total lipids. Sufficient calcium in serum 13,1% with lack of phosphorus. Ca/P ratio exceeds 1.2 times optimal value. Low alkaline phosphatase activity is typical for inadequate bone matrix formation. AST is higher on 25,1% AST and ALT parameters are both high on 25,1% and 15,4% respectively and it means liver function abnormal. There is very low vitamin B2 (42,4% from normal value) wwith sufficiently high carotene supply with vitamins A, E, C in the liver. Liver micronutrients: Zinc – 2,9 times more; Manganese – 3,1 times more with less concentration of Cobalt on 2,6 times and Selenium – 8,3 times. There are calcium 1,6 times less and phosphorus – 1,4 in tibia. Thus in the liver of chickens has focal or diffuse inflammation with blood flow disturbances and of necrosis were recorded. Macroscopic examination found that kidney are oversized, have flabby consistency with light brown color and surface look like almond nut. Microscopic examination found that Bowman's capsule of the cortex kidney is thickened, the space between the capsule and glomeruli is swollen, and the borders of the tubule epithelial cells are poorly distinguishable. The cells are swollen, enlarged and cone‐shaped into the gap make it narrow. The degree of swelling of the epithelial cells, and, consequently, the narrowing of the lumens were uneven. In some tubules, the gaps are still quite distinguishable, in others they look like gaps. There are tubules in which the gaps are almost invisible. Nuclei are not clearly expressed in all cells. The established violations in the interstitial metabolism and structural changes in the parenchymal organs (liver, kidneys) during the restoration of youth is a predisposing load that ensures the body's resistance to adverse environmental influences and loss of productive metabolism during egg production.Micromorphology of the chickens' liver. Impaired blood circulation. Hematoxylin and eosin.X 640Figure 1The architectonics of the glomerular zone of the kidney in chickens 46 days old. Hematoxylin and eosin.X 640Figure 2
Bilateral Anatomical Variation of the Upper Serratus Anterior: A Cadaveric Dissection
Authors: Etibo Kristen , Longoria Kendra , Thornton Kaitlin , Rosario Martin G. 
Published: April 2020

Anatomical variation can have clinical implications in terms of pathology, musculature surgical considerations, and biomechanical function. While many anatomical variations have been reported, it is critical to continue to expand the record when deviations are found. According to most sources, the upper serratus anterior originates at the first and second ribs and inserts at the superior angle of the scapula. Some variations in attachments of the upper serratus anterior have been described, especially regarding variations in origin at the first few ribs. (Smith, et al., 2003). However, the literature does not describe the insertion of the serratus anterior muscle as extending along the superior border of the scapula.PURPOSETo report a bilateral anatomical variation of the upper serratus anterior discovered during routine dissection on a female specimen.METHODSWhile under the supervision of the anatomist at Texas Woman’s University Dallas, Doctor of Physical Therapy students dissected an 82‐year‐old Caucasian female. The students employed techniques described in Grant’s Dissector (Detton, 2017), to expose the shoulder and neck regions of the specimen. Blunt dissection techniques were utilized to expose the area further once they discovered the anatomical variation of the upper serratus anterior. Students measured the length of the upper serratus anterior that inserted on the superior border.RESULTSThe cadaver with the serratus anterior variation, had the muscle attached to 56% of the superior border of her scapula. Comparatively, 3 other female cadavers of similar ages had significantly less attachment to the superior border‐‐an average of 27%, with a range from 16.7% to 32.6%.DISCUSSIONBased on widely accepted descriptions of the serratus anterior attaching to the superior angle, without extending extensively onto the superior border of the scapula, and in comparison to other available cadavers, this subject had a serratus anterior with a significantly larger attachment to the superior border of the scapula. The specific actions of the upper serratus anterior include stabilizing the scapula during humeral elevation, and in upward rotation and abduction of the scapula. Therefore, this variation may have functional and clinical implications for the scapulothoracic and shoulder regions.CONCLUSIONSIt appears that variations in the upper serratus anterior may not only include its rib attachments but also its scapular attachment. This may affect function including limitations in scapular rotation and shoulder elevation. Therefore, further investigation into anomalies and function of the upper serratus anterior is warranted.SA Anatomical Variation: Serratus anterior attaching to the superior angle, without extending extensively onto the superior border of the scapula.Figure 1
The vimentin rod domain targets P‐selectin to block leukocyte adhesion to inflamed endothelium
Authors: Lam Fong Wilson , Brown Cameron August , Brown Christian Valladolid , Emebo Dabel Cynthia , Cruz Miguel A. 
Published: April 2020

The acute inflammatory process begins with leukocyte adhesion to vascular endothelium and platelets. This typically beneficial process is mediated through the binding of P‐selectin glycoprotein ligand‐1 (PSGL‐1) on leukocytes to P‐selectin (P‐sel) on both inflamed endothelium and activated platelets. In pathologic conditions, this key process may lead to pathologic inflammation and contributed to secondary organ injury, as in sepsis‐induced acute liver injury and ischemia‐reperfusion injury. Therefore, the development of novel therapies to attenuate inflammation may be beneficial in select conditions. We previously described the ability of recombinant human vimentin (rhVim) to block leukocyte adhesion to endothelium and platelets and to decrease leukocytic infiltration into the lungs after endotoxemia via binding to P‐sel. The
purpose of this study was to identify the active region within rhVim that interacts with P‐selectin to further dissect the mechanism by which rhVim blocks leukocyte adhesion. Using SPOT‐peptide arrays of both full‐length vimentin and P‐sel (20‐aa/spot), we identified that rhVim binds to P‐sel primarily through its rod domain. We created a recombinant form of the rod domain (rhRod) and found that it binds to P‐sel with high affinity (10 – 41 nM), and in silico modeling suggests that it also binds to regions that interfere with PSGL‐1‐P‐sel interactions (Figure
1). In vitro experiments show that rhRod has similar efficacy to rhVim in blocking leukocyte adhesion to P‐sel/Fc coated surfaces and IL‐1β/IL‐4‐co‐stimulated endothelium (~25 – 50% reduction). Finally, using intravital microscopy in endotoxemic C57Bl/6 mice (Figure
2), rhRod co‐localizes with P‐sel in the hepatic sinusoids and treatment with rhRod decreases neutrophil adhesion to hepatic sinusoids (~25% reduction). These data suggest a potential role of rhRod in attenuating inflammation through blocking P‐selectin‐PSGL‐1 interactions.Support or Funding InformationNIH/NIGMS GM‐112806 & GM‐123261; NIH/NHLBI HL‐116524; NIH/NINDS NS‐094280; Dept. of Veterans Affairs I01 BX002551(A) Three views of the lectin/EGF‐domain of P‐selectin (PDB 1G1Q) with PSGL‐1 complexed (PDB 1G1S). The colored domains signify the regions to which rhRod bound to P‐selectin on the SPOT array (magenta: aa 151–170; cyan: aa 43–65; and blue: aa 79–116). (B) Two views of an in silico prediction model of rhRod (green) binding to P‐selectin in the region correlating to aa residues 107–129.Figure 1(A) Intravital microscopy of mouse hepatic sinusoids after endotoxin exposure show P‐selectin (red) lining the sinusoidal vessels (gray). Atto 550‐labeled rhRod (green) also localizes to the sinusoids and colocalizes with P‐selectin (yellow). Firmly adherent neutrophils (cyan) were identified as cells which moved less than 1 cell body distance over 1 minute. (B) Mice receiving rhRod had significantly fewer neutrophils per vessel area. Scale bar = 50 μm.Figure 2
A REVIEW: MALIGNANT HYPERTHERMIA‐A GENETIC DISEASE OF SODIUM CHANNEL FUNCTION
Authors: Williams Charles H. 
Published: April 2020

Malignant Hyperthermia is inherited as a dominant gene in pigs and in humans. It is characterized by a rapidly increasing body temperature up to 118°F with a metabolic rate over 10× normal, an intense peripheral vasoconstriction with blood pressures over 400 mmHg, and a fatal outcome in most cases.The MH syndrome is triggered by exercise, hauling to market, breeding activity, hot weather, and other stress factors. In humans, the cases are triggered by depolarizing muscle relaxants, Halothane anesthesia, and other halogenated anesthesia compounds. Non‐depolarizing muscle relaxants such as Pancuronium and Vecuronium are safer to use. Organon 9426 (Rocuronium) is safe to use and provides carry over protection against the development of MH.The widespread use of Sevoflurane in humans has reduced the incidence of MH to 1:550,000. Only three cases of MH have been reported in the USA during the past 30 years. They developed in WI, FL and SC.The MH susceptible pig is an outstanding animal model for research purposes and has enabled the development of new muscle relaxants and anesthesia agents that are safer for human use.The sodium channels in MH susceptible animals are leaky and allow the influx of sodium into muscle cells which must be pumped out by sodium‐potassium ATPase, thereby using ATP and generating heat.Malignant Hyperthermia is inherited as a dominant gene in pigs and in humans. It is characterized by a rapidly increasing body temperature up to 118°F with a metabolic rate over 10× normal, an intense peripheral vasoconstriction with blood pressures over 400 mmHg, and a fatal outcome in most cases.The MH syndrome is triggered by exercise, hauling to market, breeding activity, hot weather, and other stress factors. In humans, the cases are triggered by depolarizing muscle relaxants, Halothane anesthesia, and other halogenated anesthesia compounds. Non‐depolarizing muscle relaxants such as Pancuronium and Vecuronium are safer to use. Organon 9426 (Rocuronium) is safe to use and provides carry over protection against the development of MH.The widespread use of Sevoflurane in humans has reduced the incidence of MH to 1:550,000. Only three cases of MH have been reported in the USA during the past 30 years. They developed in WI, FL and SC.The MH susceptible pig is an outstanding animal model for research purposes and has enabled the development of new muscle relaxants and anesthesia agents that are safer for human use. The sodium channels in MH susceptible animals are leaky and allow the influx of sodium into muscle cells which must be pumped out by sodium‐potassium ATPase, thereby using ATP and generating heat.Support or Funding Information
Epithelial Mechanosensitive Ion Channel Piezo2 Contributes to Pressure‐Induced Epithelial Chloride Secretion in Mouse Colon
Authors: Knutson Kaitlyn Rebecca , Finholm Isabelle , Treichel Anthony J. , Whiteman Sara , Linden David R. , Grover Madhusudan , Farrugia Gianrico , Beyder Arthur 
Published: April 2020

BACKGROUNDMechanical stimulation of the gastrointestinal (GI) epithelium causes changes in epithelial ion transport by an unclear mechanism. We recently discovered the mechanosensitive ion channel Piezo2 is specifically expressed by the enteroendocrine cells (EECs) and is necessary for EEC mechanosensitivity. Further, in mouse small bowel, the pharmacologic inhibition or genetic knockout of the epithelial Piezo2 led to an inhibition of the pressure‐induced short‐circuit current (Isc) in the Ussing chamber. These results suggested that EEC Piezo2 plays a role in pressure‐induced epithelial ion transport, but the mechanism and whether these responses are region‐specific remains uknown.AIMDetermine whether epithelial Piezo2‐mediated pressure‐induced Isc is due to Clsecretion in the mouse colon.METHODSFull‐thickness mid‐colon preparations of VilCre/+::Piezo2f/f (Piezo2CKO) or Vil+/+::Piezo2f/f (Piezo2WT) mice at 0.3 cm2 were mounted in 4mL Ussing chambers with normal Krebs solution. The epithelial side was intermittently pressure‐clamped at a range of pressures (5, 10, 20, 30, 40, 60 mmHg) for 10s each. Forskolin (FSK, 10μM) was applied at the end of the experiments, and FSK response was used to normalize the pressure‐induced responses (ΔIsc/ΔIsc,FSK).RESULTSPressure applied to the mucosa of Piezo2CKO and Piezo2WTcolons caused a pressure‐dependent increase in Isc. Responses in Piezo2CKO colons were smaller than Piezo2WT at 40mmHg (0.41±0.1 vs. 0.76±0.1) and 60mmHg (0.51±0.1 vs. 0.87±0.1) (n=9–11, P<0.05, two‐way ANOVA). To determine the nature of the pressure‐induced Isc we removed Cl− from the Krebs solutions. Cl‐removal resulted in XX. We found a nearloss in the mechanically induced ΔIsc in both Piezo2WT tissues and Piezo2CKO: 5mmHg (− 88.85±0.1% and −87.44±0.1% Piezo2WT), 10mmHg (−86.46±0.1% Piezo2CKO and − 90.30±0.1% Piezo2WT), 20mmHg (−88.50±0.01% Piezo2CKO and −89.86±0.1% Piezo2WT), 40mmHg (−91.78±0.1% Piezo2CKO and −91.35±0.04% Piezo2WT), and 60mmHg (−93.93±0.04% Piezo2CKO and −92.08±0.04% Piezo2WT (n= 5–10, P <0.05, unpaired t‐test). Replacement of Cl− led to full recovery of the mechanically induced ΔIsc in both Piezo2WT and Piezo2CKO tissues when tested at 30 mmHg (Piezo2WT: 2.51±1.4 vs. 73.52±33.8, n=6 and 3, P <0.05, unpaired t‐test), (Piezo2CKO: 1.80±1.0 vs. 45.14±12.4, n=5 and 4, P <0.05, unpaired t‐test).CONCLUSIONSHydrostatic pressure applied to the colon epithelium leads to an increase in epithelial secretion that depends chloride and Piezo2 contributes to this mechanosensitive chloride secretion.Support or Funding InformationSupported by AGA RSA, NIH DK106456, NIH DK119683, NIH DK123549