bluuebunny/crossref_metadata_2025_split huggingface dataset:
Class I HDAC Differentially Regulates Blood Pressure and Renal Injury in Npr1 Gene‐targeted Male and Female Mutant Mice
Authors: Kumar Prerna , Bloodworth Meaghan , Gogulamudi Venkateswara R. , Pandey Kailash N. 
Published: April 2020

Atrial and brain natriuretic peptides (ANP, BNP) activate guanylyl cyclase/natriuretic peptide receptor‐A (GC‐A/NPRA), which plays a critical role in the regulation of blood pressure (BP) and fluid volume homeostasis, and inhibits the cell proliferation and fibrosis. Mice lacking functional Npr1 gene (coding for GC‐A/NPRA) exhibit pathophysiology of hypertension, kidney disease, and heart failure; however, the mechanisms regulating Npr1 expression are not well understood. The objective of the present study was to gain insight into the effect of class I‐specific histone deacetylase (HDAC) inhibitor, mocetinostat (MGCD0103) on Npr1 gene expression and regulation of BP and renal injury in gene‐targeted mutant mice. We utilized 16‐ to 20‐weeks old male and female Npr1 gene‐knockout haplotype (1‐copy; Npr1+/−), wild‐type (2‐copy; Npr1+/+), and gene‐duplicated heterozygous (3‐copy; Npr1++/+) mice. All mice genotypes were injected intraperitoneally with MGCD0103 (2 mg/kg) at an alternate day for 2‐weeks. The Western blot results showed that MGCD0103 significantly increased the renal NPRA protein levels in male and female mice compared with vehicle‐treated (olive oil) controls. After MGCD treatment, the renal GC‐A activity was significantly increased in both male and female mice. Male mice exhibited significantly higher systolic BP (SBP) than female mice and treatment with MGCD0103 decreased SBP in both 1‐copy male (106 ± 0.6 vs. vehicle‐treated, 129 ± 1.9; mmHg) and female (97 ± 2.2 vs. vehicle‐treated, 110 ± 2.1 mmHg, p < 0.001) mice. Renal HDAC activity was considerably higher (p < 0.05) in male mice than female mice; however, treatment with MGCD0103 attenuated (p < 0.05) HDAC activity by 40% in male and ~50% in female animals. Significantly higher urinary albumin to creatinine ratio was detected in male mice compared with female mice, which was attenuated by MGCD0103 treatment in male mice. The present results demonstrate that class I HDAC inhibitor, MGCD0103 upregulates NPRA expression in vivo, lowers SBP, and repairs renal injury differentially in male vs. female mice. These findings will have important implications for treatment of hypertension and renal injury in humans in a gender‐related manner.Support or Funding InformationThis work was supported by NIH grant HL062147.
Impaired Autoinhibition of Protein Kinase Cγ in Spinocerebellar Ataxia Type 14
Authors: Pilo Caila A. , Kunkel Maya T. , Van An-Angela N. , Kobori Evan K. , Longman Cheryl , Gorrie George , Taylor Susan S. , Newton Alexandra C. 
Published: April 2020

Spinocerebellar ataxia type 14 (SCA14) is a neurodegenerative disease caused by germline mutations in the diacylglycerol (DG)/Ca2+‐regulated protein kinase C gamma (PKCγ), leading to Purkinje cell degeneration and progressive cerebellar dysfunction. Curiously, the majority of the approximately 50 missense mutations identified in PKCγ cluster to the DG‐sensing C1A and C1B domains. Here, we use a genetically‐encoded FRET‐based C Kinase Activity Reporter (CKAR) to show that ataxia‐associated PKCγ mutants have higher basal activity in cells, and thus are less autoinhibited, than wild‐type enzyme. However, whereas reduced autoinhibition generally renders PKC sensitive to degradation, we show that mutations in the C1B domain allow translocation to membranes but protect PKCγ from phorbol ester‐induced down‐regulation. Indeed, deletion of the C1B domain prevents PKCγ down‐regulation with phorbol esters, potent ligands for the C1 domains. Strikingly, the degree of impaired autoinhibition correlates inversely with age of disease onset. Patients with the most severe mutation we examined (V138E) present with symptoms as young children, whereas symptoms in patients with the least severe mutation examined (D115Y) manifested in their 40s. To understand the structural basis of mutations outside the C1 domains, we generated a model of PKCγ using homology modeling and molecular docking. Mutations outside the C1 domains occur in regions also predicted to disrupt autoinhibition, including the pseudosubstrate, a predicted interface between the kinase and C1B domains, and the C‐terminal tail of PKCγ. Taken together, our data support a model in which SCA14 mutations enhance PKCγ activity without compromising stability. Furthermore, because many of the genetic causes of the 40+ types of SCA alter Ca2+ homeostasis, deregulated PKCγ activity may be a common cause for the disease. This raises the possibility that inhibition of PKCγ will be a potentially viable therapeutic target for SCA.Support or Funding InformationNIH T32 GM007752
Elucidation of HIV‐1 5' Leader Secondary Structure Through Novel Long Range Nuclear Magnetic Resonance (NMR) Techniques
Authors: Walker Claudia , Kan Elisabeth , Davis Faith , Catazaro Jonathan , Summers Michael 
Published: April 2020

The human immunodeficiency virus (HIV‐1) is a global pandemic that has infected approximately 37 million individuals. The high mutation rate in the HIV‐1 genome renders current treatments that target regions of the replication cycle often insufficient. Thus, structural elucidation of the 5' leader within the HIV‐1 RNA is pertinent due to the high conservation of this packaging signal, making it paramount to viral replication and a promising target for antiretroviral therapy.Structural analysis of RNA constructs below 60 nucleotides has been effective by Nuclear Magnetic Resonance (NMR) spectroscopy. However, the size of the 5' leader – approximately 300 nucleotides – has stifled structural studies by NMR due to excessive signal broadening and long instrument acquisition times. To circumvent the latter complication, site‐specific lanthanide labeling of a reporter protein with a high binding affinity to a cognate RNA loop is proposed. It is well‐known that paramagnetic lanthanide ions induce measurable pseudocontact shifts (PCS) in NMR spectra. However, binding the metal ion tag directly to the RNA is deleterious. Consequently, the spliceosomal U1A reporter protein is utilized as an intermediary between the metal ion tag and the RNA. Prior to applying the technique to the 5' leader, a 46 nucleotide Moloney Murine Leukemia Virus (MMLV) derived RNA construct was used as a model for PCS observation and NMR experiment optimization. Through the use of Heteronuclear Multiple‐Quantum Correlation experiments, PCS data was obtained in conjunction with residual dipolar couplings, resolving the quaternary structure of U1A: MMLV. Furthermore, the experiment illustrated an increase in distance measurements from 5Å to 30Å in the presence of paramagnetic lanthanides. Our novel method is currently being applied toward the elucidation of the secondary structure of the 5' leader with the U1A binding loop strategically placed in regions of interest in an effort to enhance our understanding of HIV‐1 biology and to facilitate the design of novel therapeutics in the future.Support or Funding InformationThis investigation was sponsored by NIH/NIGMS MARC U*STAR T34 HHS 00026 National Research Service Award to UMBC. This research was also supported in part by a grant to UMBC from the Howard Hughes Medical Institute through the HHMI Adaptation Project. We would like to thank the MARC U*STAR program and UMBC Summer Biomedical Training Program for the strong support.
DECREASING OF ERYTHROCYTES MEMBRANES PLASMALOGENS LEVEL AT THE DIABETES MELLITUS
Authors: Shamansurova Zulaykho , Saatov Talat , Akhrarova Nilufar 
Published: April 2020

Introduction. Plasmalogens are important part of glycerophospholipids and plays a role in the tissue development and metabolism. Changing their level shown in coronary heart diseases, peroxysomes disorders, Diabetes Mellitus (DM).Aim. Study the role of plasmalogens in the development of diabetes complication, especially in diabetic nephropathy (DN) presented interest in our study.Material and methods. In 12 healthy subjects (HS) and in 49 patients with DM2 in age 39 – 65 with disease duration from 2 month to 25 years the level of fasting blood glucose (FBG) and HbA1c, plasma triglycerides (TG) content, plasmalogens in blood serum (SP) and erythrocytes membrane (EP) were observed. The stage of DN detected by urine protein excretion level, glomerular filtration rate (GFR), blood urine nitrogen (BUN) and creatinine level. Patients according to GFR and BUN level were selected into the main 3 groups: without proteinuria (BPU), with proteinuria and normal BUN (PU), with proteinuria and higher BUN (CKD).Results. FBG and HbA1c level were significantly higher in all DM patients than HS, by 2.1, p<0.01 and in 1.7, p<0.05 respectively and was comparable between DN groups. TG level were significantly higher by 1.7 times in DM group and shown relationship with severity of DN. SP and EP level were significantly lower in DM group and showed strong linkage between GFR and BUN level and suggest about involvement of plasmalogens into development of DN.Conclusion. In patients with DM SP and EP level significantly decreased, had a strong linkage with GFR, BUN and can reflect the severity of DN.
Sustained Mentorship Promotes The Development Of Active Learning Strategies In Undergraduate Biology Classrooms: Evidence Gained From The Promoting Active Learning (PALM) Network
Authors: Wolyniak Michael J. , Wick Sue 
Published: April 2020

Here we report on the work of the Promoting Active Learning and Mentoring Network (PALM), a National Science Foundation‐supported project started in 2016 and dedicated to leveraging the expertise found in life science professional societies to provide long‐term mentorship in developing active learning techniques in the classroom. PALM mentees work one‐on‐one with mentors that are seasoned in active learning strategies and travel to their mentor’s institution to observe classes, plan modifications for their own classes, and strategize the best active learning techniques that are suited to their class’ particular needs. A monthly online journal club and regular opportunities to meet in conjunction with the Society for the Advancement of Biology Education Research (SABER) and ASCB meetings provide a dynamic teaching and learning community for mentees and mentors alike. After their mentorship, mentees are encouraged to work with their colleagues on active learning strategies and to become a PALM mentor to further the spread of effective active learning strategies to an ever‐wider audience. We have assessed the efficacy of PALM mentorship through the use of the Classroom Observation Protocol for Undergraduate STEM (COPUS) protocol (Smith et al., 2013) on pre‐mentorship and post‐mentorship videos of mentee teaching. The COPUS results have revealed a consistent shift in mentee teaching practices from a strict focus on lecture to a more diffuse combination of classroom activities in which the student is more directly active and engaged in the class and the instructor is more directly engaging students in a more personalized manner. While the specific changes in classroom practice vary between individual mentees due to the distinct needs and environments of each class, there is a consistent picture of more student‐centered classrooms that are more engaging and dynamic for both student and instructor. Overall, the COPUS data suggests that the long‐term mentoring strategies brought forth by PALM are an effective and sustainable means to promote the development of active learning techniques among undergraduate biology instructors.Support or Funding Information‐National Science Foundation Award #1624200
RNA Sequencing of Gas6 induced Preeclamptic Rat Placenta
Authors: Hirschi-Budge K. M. , Tsai K. Y.F. , Llavina S. , Hill J. , Arroyo J. , Reynolds Paul R. 
Published: April 2020

Preeclampsia (PE) is an obstetric complication that affects 2–8% of pregnancies and remains a leading cause of morbidity and mortality for both the mother and fetus. Recent research has focused on biomarkers that may indicate disease onset and models for the study of PE. One such potential biomarker is the growth arrest specific protein 6 (Gas6). Gas6 binds to the TAM family of tyrosine kinase receptors, with primary affinity for the Axl receptor. Axl‐mediated signaling is implicated in cellular proliferation and migration, and we’ve demonstrated that Axl activation by Gas6 during pregnancy induces PE in the rat, including symptoms such as increased blood pressure, proteinuria, altered trophoblast invasion, and increased inflammatory cytokines.Our objective was to elucidate changes in gene expression induced by Gas6/Axl in the developing placenta. Pregnant Holtzman Sprague Dawley rats were treated with Gas6, or Gas6 and the Axl inhibitor R428, from gestational day (GD) 7.5–17.5. Control rats received saline injections during the same period. Tissues were collected at GD 18 and snap frozen in liquid nitrogen. Placental RNA was isolated and submitted to the sequencing center at Brigham Young University. Quality filtering was performed prior to alignment, and alignment for each sample was between 98.0–98.4%. Significance was noted as an adjusted p value of < 0.05.Analysis of the RNA sequencing data identified 123 genes that were affected by Gas6 administration. Of these, 44 genes were not responsive to Axl inhibition, 29 were returned to baseline levels, and 50 were mildly to moderately responsive to Axl inhibition. Examination of enriched gene ontologies identified several impacted biological processes including cellular proliferation and apoptosis, RNA stability, lipoxin processes and metabolism, and transcription in response to stress. These initial findings highlight several options for clarifying the molecular mechanisms of PE mediated by Gas6. Further research should examine translational effects in greater detail.Support or Funding InformationThis work was supported by a grant from the Flight Attendant’s Medical Research Institute (FAMRI, PRR and JAA) and a BYU Mentoring Environment Grant (JAA and PRR).
Regulation of Expression of β‐lactamase (ampC) and Lytic Transglycosylase by a Cascade of RpoE7‐>RpoH3 Sigma Factors Regulates Ampicillin Resistance in Azospirillum brasilense Sp245
Authors: Pandey Parul , Dubey Ashutosh Prakash , Mishra Shivangi , Mishra Muktinath , Singh Vijay Shankar , Tripathi Anil Kumar 
Published: April 2020

Bacteria colonizing rhizosphere of plants face an intense competition for the nutrients present in the root exudates. This competition also involves production of antibiotics as well as of the enzymes that prevent inhibitory effects of antibiotics. Extra‐cytoplasmic function (ECF) sigma factors are known to initiate the transcriptions of genes involved in the adaptation against several stresses including cell envelope stress caused by antibiotics. Azospirillum brasilense is a plant growth promoting rhizobacterium, which colonizes the rhizosphere of a large number of non‐legume crops. The genome of A.brasilense Sp245 encodes 23 different sigma factors; one σ54 (RpoN), one σ70 (RpoD) and 21 alternative sigma factors of σ70 family. Out of 21 alternative sigma factors, 5 belong to RpoH, 10 to RpoE (ECF), and 6 to FecI type of σ factors. When the gene encoding RpoE7 was inactivated, the rpoE7::km mutant showed resistance to higher concentration of β‐lactam antibiotics compared to its parent. Inactivation of rpoE7 led to a decrease in the activity of its own promoter as well as of its divergently organized gene encoding a lytic‐transglycosylase. Transcription start site of rpoE7 was identified by 5’RACE, and studies with lacZ fusions with the promoters of rpoE7 and lytic transglycosylase genes indicated that both the promoters were regulated by RpoE7. Since β‐lactamases are known to be involved in conferring resistance to β‐lactams, we examined the expression of β‐lactamase paralogs in the parent and rpoE7::km mutant, and observed upregulation of one β‐lactamase (AmpC) in rpoE7::km compared to the wild type. The promoter activity of ampC and its regulator ampR were also increased in the rpoE7::km mutant. We also noted that RpoE7 regulated the expression of AmpC indirectly via RpoH3 sigma factor. Altogether, these data suggest that ampicillin resistance in A. brasilense is regulated by a regulatory cascade of RpoE7 and RpoH3.Support or Funding InformationParul Pandey, Ashutosh Prakash Dubey, Shivangi Mishra and Vijay Shankar Singh are Thankful to ICAR, CSIR, and UGC respectively for the research grant.
Altered Coagulation Parameters and D‐Dimer Measurements in Sepsis are useful in Scoring the Risk Stratification
Authors: Joshi Divya , Bontekoe Emily , Rondina Matthew , Middleton Elizabeth , Blair Antoinette , Siddiqui Fakiha , Hoppensteadt Debra , Fareed Jawed 
Published: April 2020

BackgroundSepsis is characterized by a simultaneous activation of inflammation and hemostasis in response to microbial infection. This systemic inflammatory response is due to the release of pro‐inflammatory cytokines, pro‐coagulants and adhesion molecules from immune cells and/or damaged endothelial tissue. Simultaneous activation of coagulation and fibrinolysis eventually leads to consumption coagulopathy and severe vascular dysfunction. This study was designed to profile coagulation parameters in a defined clinically confirmed sepsis population in conjunction with an IRB approved clinical trial.Materials & MethodsPlasma samples from septic shock patients (n=78) were collected in citrated tubes within 72 hours of ICU admission under an IRB approved protocol in conjunction with ongoing trial at the University of Utah and Veteran’s Affair FFC Health Care System VAMC. Normal controls were comprised of commercially available 25 male and 25 female samples of citrated plasma (George King Biomedical, Overland Park, Kansas City). D‐Dimer was measured using a commercially available sandwich ELISA methods. PT/INR, aPTT and fibrinogen measurements were based on clot‐based assays. All results were compiled as mean ± SD and SEM.ResultsD‐Dimer levels were markedly increase in sepsis patients (23‐fold) in comparison to control. Significant elevation of clotting parameters such as PT/INR (2.0‐fold), aPTT (2.5‐fold) and fibrinogen (2.0‐fold) was noted in the sepsis patients in comparison to controls. These differences were significant (p value ≤0.0009) for all of the parameters except fibrinogen (p value 0.4694). There was no correlation between D‐dimer and fibrinogen in the sepsis patients.Summary & ConclusionIn comparison to the control group, the sepsis patients showed wide variations in all of the parameters investigated in this study. The marked prolongation of PT and aPTT are suggestive of both the extrinsic and intrinsic pathway defects and consumption of clotting factors. The aPTT data showed wider scatter in comparison to PT data. The fibrinogen levels were also elevated and nearly 1/3 of the patients showed >1000 mg/dL levels. Marked elevation of D‐Dimer is indicative of endogenous fibrin formation and its consumption consistent with activation of secondary fibrinolysis. These results further underscore the importance of including the clotting parameters and D‐dimer in scoring the coagulopathies associated with sepsis such as disseminated intravascular coagulation.
Nitrated Fatty Acids Reduce Inflammatory Cell Recruitment in Bleomycin‐Induced Lung Injury
Authors: Krishnan Anirudh , Wilkinson Melissa L. , Gow Andrew J. 
Published: April 2020

Acute lung injury (ALI) leads to infiltration of inflammatory cells and impaired lung function. Matrix Metalloproteinase 9 (MMP9) is a zinc‐dependent endopeptidase that degrades the extra cellular matrix and increases the porosity of the lung. CD11b is a marker for macrophage activation and migration. Nitro‐oleic fatty acid (OANO2), an electrophile that reduces cardiovascular inflammation has not been examined in lung injury. OANO2 modifies cysteine residues via Michael addition altering protein function. The goal of this study were to determine if OANO2 administration would reduce MMP9 expression and cellular infiltration following ALI. An intratracheal bleomycin (ITB) ALI model was used. Two groups of 6 C57BL/6‐J mice were treated intratracheally with bleomycin (3U/kg) or PBS (50uL). Half of the animals received OANO2 (50μg) in the same instillation and 72 hours later. The lungs were collected 7 days post ITB treatment. As previously seen, mice treated with ITB lost a significant amount of body weight; however, the addition of OANO2 mitigated this loss (−2.3 +/− 0.94 vs −0.4 +/− 0.83 g). Histology revealed cellular infiltration, deposition of proteinaceous debris, and airway epithelial damage in ITB mice that was reduced with OANO2 administration. The number of CD11b+ cells (marker for migratory macrophages) and MMP9+ cells was determined by immunohistochemistry. ITB increased number of CD11b+ (69 +/− 17.1 vs 195 +/− 17.1 #cells/HPF). OANO2 treatment reduced CD11b+ cells most in ITB animals (121 +/− 17.1#cells/HPF). ITB increased MMP9+ cell numbers compared to controls (71 +/− 15.7 vs 124 +/− 15.7 #cells/HPF). However, MMP9 expression was increased with OANO2 administration in PBS animals (304 +/− 15.7; 166 +/− 15.7 #cells/HPF), though this increase was not significant in ITB mice. Increased MMP9 expression was confirmed by western blot of whole lung tissue, where both ITB OANO2 and control OANO2 animals displayed high expression. Increased MMP9 expression may result from reduced self‐degradation as a consequence of OANO2 mediated enzyme inhibition. Overall, OANO2 increased MMP9+ cells and decreased CD11b+ cell populations. These findings suggest that treatment with OANO2 may oppose ITB‐mediated pro‐inflammatory cellular activation possibly by inhibition of extracellular peptidases, making it a potential therapeutic agent for acute lung injury.Support or Funding InformationSupported by the Summer Undergraduate Research Fellowship and NIH Grants: 5R25ES020721‐09 (MPI)