bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neonatal testosterone treatment sensitizes the angiotensin II‐elicited hypertensive response in both male and female adult rats
Authors: Xue Baojian , Beltz Terry G. , Guo Fang , Johnson Alan Kim 
Published: April 2020

The sex steroid hormones, testosterone and its metabolite oestradiol, exert a profound influence on sexual differentiation and function of the neural circuits in which microglia are implicated in masculinization of brain and behavior. Our previous studies demonstrated that there were sex differences in the sensitization of hypertension, and activation of microglial cells was necessary for the induction of hypertensive response sensitization. The present study investigated whether neonatal testosterone treatment sensitizes the ANG II‐elicited hypertensive response in male and female rats when they are in adulthood. Subcutaneous testosterone (T, 1.25 mg in 25 μL corn oil) injections were performed on the dorsal surface of the pup on postnatal days 1, 3 and 5. All rats were weaned at 3 weeks of age and weighed one time per week until blood pressure experiments began. At 12 weeks of age, the T injected rats and oil injected control rats in both sexes received a subcutaneous infusion of a slow‐pressor dose of ANG II (120 ng/kg/min) for 2 weeks. Both male and female T injected rats exhibited a significant increase in body weight and had a significantly enhanced hypertensive response to the ANG II infusion when compared to control rats. These results support the hypothesis that early T‐induced masculinization in both males and females sensitizes ANG II‐induced hypertension, and suggest that masculinization disorder induced by neonatal T may predispose individuals to exacerbation of renin‐angiotensin‐elicited hypertension.Support or Funding InformationNIH HL139575
The Detection of Antibiotic Resistance Genes in Soil Samples from Rural and Urban Environments
Authors: Schuster Noah Avery 
Published: April 2020

The soil is an extremely large reservoir for microbial diversity, containing large numbers of resistant microorganisms that possess a variety of antibiotic resistance genes. Determining the composition and distribution of these genes within the soil provides information to further our understandings of antibiotic resistance evolution and development. Profiling the soil resistome can also reveal previously unknown threats to public health. We hypothesize that a larger number of resistance genes would be present in urban environments when compared to rural environments. In this study, soil was collected from twelve locations, with six locations in a rural environment and the other six in an urban environment. Soil gathered from each location was then characterized based entirely on texture and pH, followed by enrichment with tryptic soy broth containing tetracycline and incubated for three days at room temperature. Following DNA extraction from each enrichment, the presence of different tetracycline resistance genes (n = 14) was evaluated. A total of 23 tetracycline resistance genes were found to exist throughout 11 locations and the most frequently detected gene was tet(L) (30%), followed by tet(M) (26%), tetA(P) (22%), tet(O) (9%), tet(B) (9%), and tet(A) (4%). Each sequence from this study had >97% alignment when compared against known sequences of tetracycline resistance genes. In dividing sampling locations into rural and urban environments, only 12 (52%) tetracycline resistance genes were found in rural soils and 11 (48%) tetracycline resistance genes were found in urban soils. Data collected from this study does not seem to indicate any relationship between soil texture or pH versus the amount of genes present in each location. The location having the greatest number of genes (n = 5) is near local septic systems which might explain the large number of genes present. Based on function, 4 efflux and 2 ribosomal protection genes were detected; however, there is no evidence to suggest whether these genes are found more in rural or urban environments. Taken together, our data to‐date suggests that the number of tetracycline resistance genes is equally distributed in the rural and urban environments. This study hopes to continue assessing the soil resistome by analyzing DNA extracts from carbenicillin and chloramphenicol enrichments, in part with a microbial community analysis using next‐generation sequencing.Support or Funding InformationAsher Fund, DePauw University
Evidence and Inhibitor Studies of Involvement of Src and EGFR in Estrogen Receptor Induced ERK1/2 Activation in Ovarian Cancer Cell Proliferation Stimulated by Cadmium Chloride.
Authors: Steinke Kira , Huff Mary 
Published: April 2020

Cadmium Chloride, Cadmium, a naturally occurring heavy metal, is a toxic industrial and environmental pollutant commonly found in both ground and industrial waste water. Studies have shown that cadmium is also a metalloestrogen that can bind to the human estrogen receptor (ER), stimulate DNA synthesis, and increases cellular proliferation in ovarian cancer cell lines. Furthermore, it has been shown that cadmium stimulates ERK1/2 activity in an ER‐dependent manner. To determine if cadmium stimulates proliferation in ovarian cancer cell lines, OVCAR3 and SKOV3 cells were treated with varying concentrations cadmium chloride, and growth was measured by a cell proliferation assay. The results support that cadmium induces cellular proliferation at environmentally relevant concentrations (0.001–1.0 uM) within 24 hours of treatment. It was also observed that treatment with 1 nM of cadmium for 5 minutes activates ERK1/2 phosphorylation. We are now in the process of performing inhibitor studies to determine if Src and EGFR are also involved in ER dependent cadmium‐stimulated cellular proliferation in ovarian cancer cells.Support or Funding InformationBellarmine University, Student Government Association
PIP2 Binds Multiple Cationic Sites of ENaC
Authors: Archer Crystal R. , Carver Chase M. , Enslow Benjamin T. , Stockand James D. 
Published: April 2020

Phosphatidylinositol 4,5‐bisphosphate (PIP2) regulates the activity and gating of many different ion channels to include the epithelial Na+ channel (ENaC). The negatively charged phosphoryl groups of the PIP2 headgroup are expected to interact with cationic residues of the cytoplasmic tails of ENaC. To date, understanding of the biophysical interaction between ENaC and PIP2 has been limited to indirect functional assays. Consequently, the precise binding sites and their affinities have not been defined. We began this study using secondary structure prediction tools to show that helical conformation that may cluster the cationic residues of ENaC cytoplasmic tails. Such clustering would form an electropositive region attractive to the PIP2 headgroup. We used a CIBN/CRY2‐5‐ptase optogenetic dimerization system in HEK cells to show that the C5 phosphoryl group of PIP2 plays a role in regulating ENaC activity. To more precisely define the PIP2 binding sites of ENaC, we designed short peptides corresponding to the cationic clusters of ENaC. Steady state intrinsic fluorescence ligand binding data showed that PIP2 interacted with the extreme amino terminus of the β‐ENaC subunit (βN1 peptide) and the amino and carboxy termini of γ‐ENaC nearest the transmembrane domains (γN2 and γC peptides, respectively), but no significant changes were observed with the other peptides or mutant controls. Microscale thermophoresis data revealed that the βN1 and γN2 sites has the strongest interaction with the PIP2 headgroup with Kd ~5.2 and 15 μM, respectively, whereas the γC site interacts with much weaker affinity. Again, no interaction was observed between PIP2 and the other peptides and mutant controls. These results are consistent with PIP2 binding ENaC to stabilize the open conformation of channel via moderate and weak electrostatic interactions within the cytoplasmic termini.Support or Funding InformationThis work is supported by NIH/HHLBI T32 HL07446 to CRA and JDS, and NIH/NIDDK R01 DK987460 and DK117909 to JDS.
Kidney mitochondrial redox state 3 days following uninephrectomy in rats determined by cyro‐fluorescence optical imaging.
Authors: Cowley Allen W. , Yang Chun , Shimada Satoshi , Zheleznova Nadya , Mehrvar Shima , Foomani Farnaz , Mostaghimi Soudeh , Ranji Mahsa 
Published: April 2020

Uninephrectomy (UNX) is known to result in structural and metabolic changes to the remaining kidney although it is uncertain if this results in changes in the mitochondrial redox state and how soon such changes may occur. A custom designed fluorescence cryo‐imaging technique was used to quantitatively assess the effect of UNX on the mitochondrial bioenergetics of the remaining kidney. Responses were obtained in kidneys of normal Sprague Dawley (SD) rats, Dahl salt‐ sensitive (SS) rats, and SS rats in which the gene for NADPH‐oxidase isoform 4 (NOX4) was knocked out (SSNOX4−/−). Kidneys were snap frozen and the intrinsic fluorescence of two coenzymes, Nicotinamide Adenine Dinucleotide (NADH) and oxidized Flavin Adenine Dinucleotide (FAD) were optically acquired and 3D images created to characterize the NADH/FAD redox ratios (RR) of each kidney 3 days following UNX. Each of the strains exhibited similar increases of kidney weights averaging 17% after 3 days of UNX. It was especially notable that following UNX, SD and SSNOX4−/− rats exhibited global reductions of the RR ratios (P<0.05) with a similar tendency observed in SS rats (P<0.08) indicative of increased reactive oxygen species (ROS) production. The unexpected reduction of the mitochondrial RR in the remaining kidneys of SSNOX4−/− rats indicates that mechanisms independent of H2O2 produced from NOX4 may be largely response for this global increase of renal oxidative stress. We propose that enhanced mitochondrial bioenergetics due to increased tubular workload of the remaining kidney becomes the dominant source of increased ROS following UNX and could represent an important hypertrophic signaling mechanism.Support or Funding InformationHL‐116264
Identification of Molecules by Spectral Imaging
Authors: Alshammari Qamar Aqeel , Pala Rajasekhar , Katzir Nir , Nauli Surya M. 
Published: April 2020

Spectral imaging (SI) is a powerful technique which primarily uses the wavelength to identify and quantify the molecules precisely with their exact location. It facilitates the identification of chemical substances and studying their properties through analyzing the way they interact with light. The study of light interaction with elements is called spectroscopy; spectroscopy examines how light behaves in the target and recognizes materials based on their spectral signatures. These spectromicroscopic methods provide a quantitative analysis of a biological sample and its substructures with a depth of analysis that no other analytical technique can determine from the same sample. Therefore, we hypothesize that identifying the exact location and quantity of molecules present in the given biological (LLCPK‐1 cells) samples can be done by using a SI system. In this study, we first measured the exact UV‐Vis absorption and fluorescence spectra of our test substances of Rhodamine 6G and Doxorubicin, independently. After that, we used the Quadrupole Time‐of‐Flight Liquid Chromatography/ Mass Spectrometer (Q‐TOF LC/MS ) system to quantify the maximum and minimum detectable concentrations of Rhodamine 6G and Doxorubicin by checking the molecular weight of mass‐spectra. In addition, we used High Performance Liquid Chromatography (HPLC) system to quantify the same chemicals based on their UV spectrum at 218 nm. Finally, we used SI microscope to determine the exact amount and location of the molecules present in the cells. For Rhodamine 6G and doxorubicin, we started with the minimum detectable concentrations by Q‐TOF and consider it as a maximum limit for the SI. Using the SI we were able to detect the specific location of Rhodamine 6G was in the cytoplasm and Doxorubicin in the nucleoplasm. In conclusion, SI was able to detect much more lower concentrations of Rhodamine 6G, in the cytoplasm, and Doxorubicin, in the nucleoplasm, based on the spectrum in comparison to Q‐TOF LC/MS.Support or Funding InformationSaudi Arabian Cultural Mission
Functional Connectivity of Prelimic Prefrontal Cortex and Rostral Ventromedial Medulla for Descending Pain Modulation
Authors: Smith Angela , Flohrschutz Austin , Riegel Arthur , Largent-Milnes Tally , Vanderah Todd 
Published: April 2020

Chronic pain is a health condition that can seriously impact quality of life. To develop better treatments for chronic pain, a better understanding of the mechanisms behind pain perception is needed. Current models of descending pain modulation show the prelimbic prefrontal cortex (plPFC; area 32V) synapsing onto cells within the periaqueductal grey (PAG) which then synapses onto the rostral ventromedial medulla (RVM). The RVM contains ‘on’ and ‘off’ cells which respond to pain stimuli by respectively activating or inhibiting pain perception. We aim to functionally characterize a hypothesized direct projection from the plPFC to the RVM first described in 1997. Anterograde tracing was performed in the plPFC to determine the anatomical connectivity of the two regions. Similarly, we transfected the plPFC with hM4Dq DREADDs fused with mCherry under the control of CaMKIIa promoters to label glutamatergic Layer 5 projection neurons and determine functional connectivity of the purported circuit in animals with a spared nerve injury (SNI). These receptors are inert unless acted on by a designer drug, such as CNO. We used reverse microdialysis to locally deliver CNO into the RVM and activate DREADDs on neurons with direct projections from the plPFC while collecting released neurotransmitter and performing reflexive pain behaviors assays. This strategy allowed for avoidance of systemic/off target activation of plPFC neurons projecting to a different CNS region. Anatomical and microanalysis pilot data indicate a direct connection from plPFC to the RVM.Support or Funding InformationFunding provided by the University of Arizona Department of Medicinal Pharmacology, University of Arizona Undergraduate Biological Research Program, and the ASPET Summer Undergraduate Research Fellows Program.
Antioxidant Capacity and Antiproliferative Activity of Morchella esculenta in Human Ovarian Adenocarcinoma Cells
Authors: Scales Karissa , Schlacks Alexander , Parr Tyler , Soendergaard Mette 
Published: April 2020

Morchella esculenta is a coveted wild edible mushroom that has shown antiproliferative activity in various cell lines. However, little is known about the effects of M. esculenta on the cell viability of ovarian cancer. Here, the antiproliferative activity of water, ethanol, and methanol extracts of “grey” and “yellow” M. esculenta fruiting bodies on human ovarian adenocarcinoma cells were investigated. Further, the antioxidant capacity and phenolic content of the mushrooms were evaluated.Wild “grey” and “yellow” M. esculenta fruiting bodies were harvested in Illinois, USA in April 2019, immediately freeze‐dried, and ground into powders. For antioxidant assays, M. esculenta powder was extracted using 1% HCl in 90% aqueous methanol for 2 h. The antioxidant capacity was evaluated using 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) assays. Both assays revealed significantly (p<0.0001) higher antioxidant capacity (trolox equivalents; TE; μmol/g dry weight) of “grey” compared to “yellow” mushrooms. Specifically, the DPPH assay showed antioxidant activities of 5.35±0.152 and 6.99±0.0190 TE (mean±SEM) for “yellow” and “grey” M. esculenta, respectively. The antioxidant capacity as determined by the ABTS assays was 3.87±0.113 and 9.78±0.112 TE (mean±SEM) for “yellow” and “grey” mushrooms, respectively. The Folin‐Ciocalteu method was used to determine the total phenolic content (gallic acid equivalents; GAE; μmol/g dry weight). In correlation to the DPPH and ABTS results, the phenolic content was significantly higher (p<0.0001) for “grey” compared to “yellow” mushrooms. The phenolic content was determined to be 9.90±0.098 and 12.02±0.316 GAE (mean±SEM), respectively.For analysis of antiproliferative activity, mushroom powders were extracted thrice for 24 h using water, ethanol, or methanol. Extracts were then air‐dried overnight at 50°C and resuspended in dimethyl sulfoxide (DMSO) to 20 mg/mL. Human ovarian adenocarcinoma (SKOV‐3) cells were then treated with 0.2 mg/mL mushroom extract, 100 μM paclitaxel, or DMSO (vehicle) for 48 h at 37°C, 5% CO2. Cell viability was determined using a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) reduction assay by measuring the absorption of formed formazan at 570 nm. Water and ethanol extracts from both “yellow” and “grey” M. esculenta resulted in significantly reduced cell viability (p<0.05) compared to DMSO. However, methanol extracts failed to show any effect on cell viability. Based on these results, water and ethanol extracts from “yellow” and “grey” M. esculenta significantly lower the cell viability of ovarian adenocarcinoma cells. These mushrooms may, therefore, exhibit antiproliferative potential against ovarian cancer cells.
Role of Histone Deacetylases 5 (HDAC5) in Ang II‐Induced the Early Growth Response Protein‐1 (Egr‐1) Expression and Vascular Hypertrophy
Authors: Truong Vanessa , Jain Ashish , Anand-Srivastava Madhu , Srivastava Ashok 
Published: April 2020

Angiotensin II (Ang II), a key vasoactive peptide, is known to play an important role in the pathophysiology of vascular diseases. A heightened activation of Ang II‐induced signaling pathways that promote proliferation, hypertrophy and migration of vascular smooth muscle cells (VSMCs) has been suggested to contribute to vascular dysfunctions. We have shown earlier that Ang II enhances the expression of early growth response protein‐1 (Egr‐1), a zinc transcription factor, which is upregulated in animal models of vascular diseases. Histone deacetylases (HDACs) remove acetyl groups of lysine residues from histone and non‐histone proteins and elevated activation of class IIa HDACs is reported in several vascular pathologies. Recent studies have demonstrated that the phosphorylation and the nuclear export of HDAC5, a member of class IIa HDACs, is associated with vascular remodeling through their ability to interact with transcription factors linked to cell proliferation, migration and hypertrophy. However, the involvement of HDAC5 in Ang II‐induced Egr‐1 expression remain unexplored. Here, we showed that the specific pharmacological inhibition of class IIa HDACs by MC1568 or TMP‐195 as well as siRNA‐mediated silencing of HDAC5 attenuated Ang II induced Egr‐1 expression in VSMCs. In addition, the blockade of the nuclear export of HDAC5 by using nuclear transport inhibitors leptomycin B or KPT‐330 decreased Ang II‐induced Egr‐1 expression. We also demonstrated that siRNA‐induced silencing of HDAC5 or Egr‐1 prevented VSMC hypertrophy in response to Ang II stimulation. In summary, our results show that Ang II‐induced Egr‐1 expression is regulated by the activation of HDAC5 and Ang II‐induced cell hypertrophy is mediated by HDAC5 and Egr‐1.Support or Funding InformationSupported by the Canadian Institutes of Health Research
Immunohistochemical expression of β‐Catenin associated to bone strain changes in the subchondral bone of mandibular condyle – study in Wistar rats with dental premature contact
Authors: Freire Alexandre Rodrigues , Rossi Ana Cláudia , Okamoto Roberta , Ferreira Beatriz Carmona , Prado Felippe Bevilacqua 
Published: April 2020

From the biomechanical concepts regarding the masticatory load distribution, the mandible has a load flow pattern from the teeth to the temporomandibular joint (TMJ). Studies have presented morphological changes in the temporomandibular joint after dental occlusal alterations, such as the premature contacts. Thus, it is important to figure out the bone mechanobiology of the mandibular condyle, involving the relation mechanical stimuli x biological response. The aim of this work was to perform an immunohistochemical analysis of β‐catenin expression associated to a computational simulation by finite element analysis (FEA) to evaluate the relation between the osteocyte mechanosensation and the bone strain. This relation was evaluated in rats with occlusal changes by premature contact. Thirty male rats (Rattus norvegicus albinus), Wistar lineage and 2 months old were used in this study. The animals were divided into two groups: control group (N = 10) and experimental group (N = 20). In the experimental group, premature contact was created by a stainless‐steel fragment, with 0.7 mm thickness, cemented on the first upper molar of all animals. Then, these animals were subdivided into two groups, according to the period with premature contact of 7 days (7D) and 14 days (14D). After euthanasia, the immunohistochemical analysis was performed by indirect immunoperoxidase method for β‐catenin expression, using diaminobenzidine for staining and hematoxylin for counter stain. From the head microCT scans of control and experimental groups, finite element models were created and the FEA was performed for both groups, considering the presence of premature contact in experimental group. In all analyses, the subchondral bone of condyle was analyzed in three regions (medial, central and lateral). The immunohistochemical analysis presented higher expression of β‐catenin in the lateral region in all groups as well as the bone strain in FEA. The experimental group presented higher β‐catenin expression 7D compared to control group. At 14D the β‐catenin expression the bone strain was similar to the control group. Thus, in conclusion, the β‐catenin expression can be associated to the strain sensitivity in the subchondral bone of the mandibular condyle. The premature contact increased the bone strain and the expression of β‐catenin in the subchondral bone.Support or Funding InformationBrazilian Coordination for the Improvement of Higher Education Personnel (PNPD/CAPES).
Functional Compensation Precedes Recovery of Tissue Mass Following Acute Liver Injury
Authors: Walesky Chad , Kolb Kellie , Winston Carolyn , Henderson Jake , Kruft Benjamin , Fleming Ira , Ko Sungjin , Monga Satdarshan P. , Mueller Florian , Apte Udayan , Shalek Alex , Goessling Wolfram 
Published: April 2020

The liver plays a central role in metabolism, protein synthesis and detoxification. It possesses unique regenerative capacity upon injury. While many factors regulating proliferation during liver repair have been identified, the mechanisms by which the injured liver maintains vital functions prior to tissue recovery are unknown. Here, we identify a new phase of functional compensation following acute liver injury that occurs prior to cellular proliferation. By coupling single‐cell RNA‐seq with in situ transcriptional analyses in two independent murine liver injury models, we discover adaptive reprogramming to ensure expression of both injury response and core liver function genes dependent on macrophage‐derived WNT/β‐catenin signaling. Interestingly, transcriptional compensation is most prominent in non‐proliferating cells, clearly delineating two separate phases of liver recovery. Overall, our work describes a new mechanism by which the liver maintains essential physiological functions prior to cellular reconstitution and characterizes macrophage‐derived WNT signals required for this compensation.Support or Funding InformationC.W. is supported by F32DK111151, the Cholangiocarcinoma Foundation Research Fellowship, and the American Liver Foundation Charles Trey, MD Memorial Post‐doctoral Research Fellowship. W.G. is supported by R01DK090311, R01DK105198, R24OD017870, and the Claudia Adams Barr Program for Excellence in Cancer Research. W.G. is a Pew Scholar in Biomedical Sciences. A.K.S. was supported, in part, by the Searle Scholars Program, the Beckman Young Investigator Program, the Pew‐Stewart Scholars Program for Cancer Research, a Sloan Fellowship in Chemistry, the NIH (1DP2GM1194192RM1HG0061931R01DA046277, and the Bill and Melinda Gates Foundation. U.A. is supported by R01DK098414.