bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Neem Extracts on Formed Biofilm of Pseudomonas aeruginosa
Authors: Kaverimanian Vishnu , Heuertz Rita M. 
Published: April 2020

Over time, bacteria have been developing increasing resistance to commonly used antibiotics in medicine. To compound this issue, certain surface adherent bacteria can undergo a community response to form biofilms, a protective layer consisting of extracellular polymeric matrix that enhances resistance to antibiotics and host defenses. Biofilm association can confer bacteria with up to 1,000 times more antibiotic‐resistance. According to the NIH, up to 65% of all human infections and 80% of chronic infections are biofilm‐associated. P. aeruginosa, a common nosocomial pathogen exhibiting antibiotic resistance, is a known biofilm producer. Due to the prevalence of biofilms and their role in the antibiotic resistance of common nosocomial pathogens, it is important to look for antibiofilm agents. One understudied source of antibiofilm agents is plant derived compounds (phytochemicals). Of note, neem leaf extracts have been shown to inhibit biofilm formation in scientific literature. The purpose of my project was to identify the effect of neem extracts (bark, fresh leaf, dried leaf) on eradicating pre‐formed biofilm of P. aeruginosa (ATCC 27853) via a microplate biofilm quantitative assay. Bacteria were first inoculated into tryptic soy broth (TSB) and incubated (37°C, overnight). Afterwards, bacteria were density standardized (A590=0.2), transferred to microplates (polystyrene U‐bottom 24‐well), and incubated (37°C, overnight) to form biofilm. To isolate and treat the pre‐formed biofilm, microplates were washed (3 times, TSB), TSB was replaced, neem extracts were placed in the appropriate wells (a final concentration of 10% for each neem extract), and plates were incubated (37°C, 5 days). Afterwards, biofilm associated bacteria were killed with glutaraldehyde (10%) and microplates were washed again (3 times, dH2O). Subsequently, crystal violet (0.1% aqueous) was added to stain the biofilm. Excess dye was removed by washing (3 times, dH2O) and stained biofilm was solubilized (8% acetic acid). Solubilized biofilm was quantitated by A590 readings. Results indicated that all the neem extracts had no significant (p<0.05) effect on biofilm eradication for P. aeruginosa 27853. This suggests that neem extracts may be more effective on developing biofilm rather than formed biofilm. Further strains will be tested for neem effect on biofilm eradication.Support or Funding InformationFinancial support for VK was received from the DeNardo Education and Research Foundation.
High‐fat diet increases p62 and mTOR phosphorylation in diabetic eNOS deficient mice
Authors: George Jasmine , Chen Xiaoming , Zhao Xueying 
Published: April 2020

Diabetic nephropathy is the leading cause of chronic kidney disease and end stage renal disease. Emerging evidence has suggested that dysregulated renal autophagy may contribute to both glomerular and tubulointerstitial damage under diabetic conditions. This study was designed to elucidate the effect of a high‐fat diet on autophagy in eNOS deficient animals with and without hyperglycemia. Six‐week old wild‐type (WT) C57BL/6J and eNOS deficient mice were placed on either a normal diet (ND, 4.5% from fat) or high‐fat diet (HFD, 45% from fat). Two weeks later, half of them were rendered diabetic with alloxan. Animals were maintained on ND or HFD for 5 months post diabetes induction. Kidney damage and autophagy activity were evaluated in diabetic WT and eNOS deficient mice. HFD and diabetes induction in eNOS deficient mice resulted in significantly increased urinary albumin to creatinine ratio, matrix metalloproteinase‐9, and kidney injury molecule‐1. Western blot further indicated a significant increase in autophagy substrate p62 in kidney cortex of diabetic eNOS deficient mice fed with HFD, whereas there was no notable change in p62 expression in WT mice. In addition, mammalian target of rapamycin (mTOR) phosphorylation was increased in diabetic eNOS deficient mice. Taken together, our results suggest that HFD may impair renal autophagy by activating mTOR in diabetic eNOS deficient mice.Support or Funding InformationNIH (SC1DK112151), NIH/NCRR /RCMI (8G12MD007602) and (8U54MD007588), and R25GM058268
Effects of Ambient Temperature and Social Housing on Locomotor Activity, Thermoregulation, and Weight Loss Following “Binge” Administration of Structurally‐Related Abused Psychostimulants
Authors: Russell Lauren , Hyatt William , Urquhart Kyle , Fantegrossi William 
Published: April 2020

3,4‐methylenedioxypyrovalerone (MDPV) and α‐pyrrolidinopentiophenone (α‐PVP) are common active constituents of abused “bath salts” products. The structures of these synthetic cathinone analogues differ only in that MDPV contains a methylenedioxy ring. Additionally, MDPV is structurally similar to the amphetamine analogue 3,4‐methylenedioxymethamphetamine (MDMA), which differs fromthe structure of methamphetamine (METH) only by the addition of a methylenedioxy ring. MDPV, α‐PVP, MDMA, and METH are four psychostimulants often used interchangeably by humans in crowded, hot environments. The effects of MDMA are known to be dramatically impacted by crowding and ambient temperature and because of the structural similarities and use patterns in humans, it was of interest to investigate the modulatory effects of such environmental variables on the in vivo pharmacology of the four compounds in mice. In the present studies, adult male NIH Swiss mice were housed singly or in groups of 3 or 6, at 20°C, then administered a “binge” regimen of 1.0, 3.0, or 10.0 mg/kg MDPV, α‐PVP, MDMA or METH, or repeated saline, via intraperitoneal injection, every 2 hours for a total of 4 injections. Locomotor activity and core temperature were continuously collected throughout drug administration, continuing 2 hours after the 4th injection. The lowest dose per social housing condition different from saline was repeated at an elevated temperature of 28°C. Doses were limited by lethal effects which were potentiated both by the increased ambient temperature and social housing conditions. Increased ambient temperature resulted in increased locomotor activity for all compounds, and an increased hyperthermic effect occurred in animals administered MDPV or MDMA, both methylenedioxy ring containing compounds. No pronounced differences regarding body weight and core temperature as a function of social housing were observed. Brain regions (including hippocampus and striatum) were collected for neurochemical analysis 10 days after the last injection. Regions from one hemisphere will be processed and analyzed for tissue content of monoamines and their primary metabolites by HPLC, while samples from the other hemisphere will be utilized to determine GLUT2 concentration via western blotting. These studies suggest that the addition of specific moieties, in this case a methylenedioxy ring, to structurally similiar cathinones and amphetamines can result in same effect compared to the parent compound.Support or Funding InformationThese studies supported by DA039195, DA022981 and the UAMS Center for Translational Neuroscience.
Nicotinamide mononucleotide (NMN) supplementation rescues cerebromicrovascular endothelial function and neurovascular coupling responses and improves cognitive function in aged mice.
Authors: Tarantini Stefano , Valcarcel-Ares Marta Noa , Toth Peter , Yabluchanskiy Andriy , Kiss Tamas , Ballabh Praveen , Farkas Eszter , Baur Joseph , Sinclair David , Csiszar Anna , Ungvari Zoltan 
Published: April 2020

Adjustment of cerebral blood flow (CBF) to neuronal activity via neurovascular coupling (NVC) has an essential role in maintenance of healthy cognitive function. In aging increased oxidative stress and cerebromicrovascular endothelial dysfunction impair NVC, contributing to cognitive decline. There is increasing evidence showing that a decrease in NAD+ availability with age plays a critical role in a range of age‐related cellular impairments but its role in impaired NVC responses remains unexplored. The present study was designed to test the hypothesis that restoring NAD+ concentration may exert beneficial effects on NVC responses in aging. To test this hypothesis 24‐month‐old C57BL/6 mice were treated with nicotinamide mononucleotide (NMN), a key NAD+ intermediate, for 2 weeks. NVC was assessed by measuring CBF responses (laser Doppler flowmetry) evoked by contralateral whisker stimulation. We found that NVC responses were significantly impaired in aged mice. NMN supplementation rescued NVC responses by increasing endothelial NO‐mediated vasodilation, which was associated with significantly improved spatial working memory and gait coordination. These findings are paralleled by the sirtuin‐dependent protective effects of NMN on mitochondrial production of reactive oxygen species and mitochondrial bioenergetics in cultured cerebromicrovascular endothelial cells derived from aged animals. Thus, a decrease in NAD+ availability contributes to age‐related cerebromicrovascular dysfunction, exacerbating cognitive decline. The cerebromicrovascular protective effects of NMN highlight the preventive and therapeutic potential of NAD+ intermediates as effective interventions in patients at risk for vascular cognitive impairment (VCI).Support or Funding InformationThis work was supported by grants the Oklahoma Center for the Advancement of Science and Technology, the Oklahoma Shared Clinical and Translational Resources (OSCTR) program funded by the National Institute of General Medical Sciences (GM104938), the NIA‐supported Geroscience Training Program in Oklahoma (T32AG052363), the Oklahoma Nathan Shock Center (P30AG050911), the Cellular and Molecular GeroScience CoBRE (1P20GM125528, sub#5337), and the NIGMS supported Center of Biomedical Research Excellence (CoBRE).
Myology of the Reptilia
Authors: Holliday Casey M , Wilken Alec , Verhulst Conner , Bales London , Sellers Kaleb , Lessner Emily , Middleton Kevin 
Published: April 2020

Sauropsid vertebrates (lepidosaurs, turtles, crocodilians and birds) have evolved a diversity of head shapes and feeding behaviors during their history. A key to understanding this great radiation of reptiles is the physiology of the jaw musculature which powers the feeding apparatus. However we still know little about jaw muscle mechanics within lineages of reptiles or how this complicated musculoskeletal system has evolved to employ a variety of behaviors. New imaging and computational methods are now enabling an extraordinary view into the 3D anatomy and biomechanics of reptiles and other vertebrates. Here we illustrate several approaches to analyzing jaw muscle morphology and architecture using contrast imaging, 3D fiber tracking, biomechanical analysis, and data visualization methods that offer enormous potential for exploring the anatomy, function and evolution of jaw muscles. We first illustrate basic workflow of 3D jaw muscle visualization, morphometrics and interpretation using crocodilian jaw muscle anatomy. Second, we show how homologous jaw muscle bellies evolve among lineages of different reptiles and birds to elicit different functional demands. Third, we show how the 3D architecture of hard‐to‐dissect, small, deep protractor muscles correlate with different types of cranial kinesis among a sample of lizards and birds. These new imaging and analytical approaches offer incredible potential for the quantification of soft tissue morphology like muscle architecture and their applications to comparative biomechanics, ecomorphology, and evolutionary biology should prove remarkable.Support or Funding InformationNSF IOS 1457319, NSF EAR 1631684, University of Missouri Life Sciences Fellowship, University of Missouri Integrative Anatomy Fellowship, IMSD‐Express
Identification of Putative RNA Methyltransferases in Trypanosoma brucei
Authors: Williams Laura G , Smith Xiane , Taber Cassandra C , Ellis Lauren , Militello Kevin T 
Published: April 2020

Human African Trypanosomiasis (African sleeping sickness) is a potentially fatal disease in sub‐Saharan Africa that is caused by the protozoan parasite Trypanosoma brucei. This vector‐borne disease afflicts tens of thousands of individuals every year, and without a vaccine, it is a prominent threat to the health and safety of many African communities. RNA methylation may be an important mechanism for the regulation of gene expression in T. brucei, as this organism lacks regulatory DNA sequences. The presence of 5‐methylcytosine has been detected in the RNA of T. brucei through sodium bisulfite sequencing. Seven putative RNA methyltransferases were identified in the genome of the parasite via bioinformatics and were named TbCRMTs (
T.
brucei
cytosine
RNA
methyl
transferases). The levels of TbCRMT RNA in both procyclic and bloodstream forms were measured via RT‐qPCR and demonstrated the expression of all seven genes in both life cycle stages. TbCRMT4 is 103
kDa, and contains the conserved SAM binding domain and catalytic cysteines found
inbona fide RNA methyltransferases. TbCRMT4 is required for maximum parasite growth as determined by RNAi knockdowns. In an attempt to determine whether TbCRMT4 functions as an RNA methyltransferase, the gene was expressed in E. coli with an N‐terminal 6x‐His tag. SDS‐PAGE results suggested that the full‐length protein was fragmented due to the presence of multiple bands. In light of this, the 6x‐His tag was moved to the C‐terminus and fragments of the gene containing putative SAM‐binding and catalytic sites were then amplified by PCR. The PCR products were then inserted into the pET101/D vector and transformed into competent E. coli. Expression of each construct in E. coli was induced under different conditions, with varying degrees of success. The construct which
contains amino acids 87‐852, when expressed at 37°C, is most reliably produced, as confirmed via SDS‐PAGE/Coomassie blue staining and Western Blot analysis. Isolation of this protein was attempted using denaturing protocols and His‐affinity chromatography. Solubility of the protein has proven difficult, thus optimization of protein isolation is still ongoing. Ultimately, we plan to isolate the TbCRMT4 87‐852 protein and conduct a series of methyltransferase assays, which will determine whether or not this protein catalyzes RNA methylation. By determining if TbCRMT4 functions as an RNA methyltransferase, we can further understand the T. brucei epitranscriptome.Support or Funding InformationNIH grant 1R15AI133428‐01 to KTM
Characterization of the Dual Pathways for Bacillus halodurans Asparaginyl‐tRNA Formation
Authors: Bile Jon Matthew , Sheppard Kelly 
Published: April 2020

Protein synthesis requires the attachment of an amino acid to its cognate transfer RNA (tRNA). Two distinct pathways for attaching asparagine (Asn) to tRNAAsn are known: the direct pathway, in which Asn is directly attached to tRNAAsn by the enzyme AsnRS; and the indirect pathway, in which Asn is attached to tRNA by a non‐discriminating AspRS and GatCAB. Bacillus halodurans uses both routes for Asn‐tRNAAsn formation. To better understand why B. halodurans possesses both pathways, we are purifying its AsnRS and AspRS in order to test and compare their activities under different chemical conditions such as reactive oxygen species and pH. This research will provide insight into how B. halodurans has adapted to survive in different environmental conditions.Support or Funding InformationNational Science Foundation (MCB‐1615770)
Quantifying the Stability of Human Cytomegalovirus RNA5.0
Authors: Tsingine Hannah P. 
Published: April 2020

HCMV is the largest of the human herpesviruses containing a DNA genome of approximately 240 kilobases encoding 225 open reading frames (ORFs). Its large and complex genome also encodes long noncoding RNAs (lncRNA) which are known for not being translated, but rather function as RNA. In HCMV, one of the lncRNA, known as RNA5.0, is a stable intron that is believed to play an important role in viral pathogenesis. Recent studies have shown that RNA5.0 is a nuclear localized stable intron that is transcribed by RNA polymerase II, and that it's also enriched with high adenine and thymine residues. The true role of RNA5.0 is still unknown. It could activate transcription, regulate gene silencing, or play a role in HCMV latency. To begin to understand why an intron that should be degraded within minutes is stable for hours, we are developing an assay to quantify the stability of these unusual lncRNA using transfected cultured 293T cells.Support or Funding InformationNIH MARC U*STAR recipient T34GM092711
Angiotensin‐(1‐7) Improves Integrated Cardiometabolic Function in Aged Mice
Authors: Miller Amanda Joy , Bingaman Sarah , Mehay Daren , Medina Daniela , Arnold Amy Christine 
Published: April 2020

BackgroundAging is associated with impaired cardiometabolic function including insulin resistance, elevated blood pressure, and cardiac hypertrophy. Angiotensin (Ang)‐(1‐7) is a beneficial hormone of the renin‐angiotensin system that improves control of blood pressure and glucose homeostasis in models of cardiovascular and metabolic diseases in young mice. To our knowledge, no studies have evaluated the importance of Ang‐(1‐7) to integrated cardiometabolic function in aging. We hypothesized that Ang‐(1‐7) would improve insulin sensitivity as well as decrease blood pressure and cardiac hypertrophy in healthy aged mice.MethodsSixteen month‐old male C57BL/6J mice received 6‐week Ang‐(1‐7) [400 ng/kg/min, n=9] or saline infusion (n=9) via subcutaneous osmotic mini‐pumps. All mice were fed standard chow diet (18% kcal from fat). During the last week of treatment, intraperitoneal insulin and glucose tolerance tests were performed and blood pressure was measured via a carotid artery catheter. Heart weight was measured at the end of experiments. Gene expression of tyrosine hydroxylase (index of sympathetic activity), inflammatory markers (IL10, IL1β, IL6, TNFα) and Ang II AT1 and Ang‐(1‐7) Mas receptors were measured in myocardial tissue by quantitative real‐time PCR and quantified with 2‐ΔΔCT methods.ResultsWe found that Ang‐(1‐7) decreased systolic [Ang‐(1‐7): 105±2 mmHg, saline: 116±4 mmHg, P=0.031], diastolic [Ang‐(1‐7): 77±2 mmHg, saline: 83±2 mmHg, P=0.041], and mean [Ang‐(1‐7): 86±2 mmHg, saline: 94±3 mmHg, P = 0.023] blood pressure. Furthermore, Ang‐(1‐7) reduced heart weight [Ang‐(1‐7): 0.18±0.01 g; saline: 0.21±0.01 g, P=0.042]. The improvement in cardiac hypertrophy with Ang‐(1‐7) was associated with decreased tyrosine hydroxylase gene expression in the heart [Ang‐(1‐7): 0.4±0.1, saline: 0.9±0.1, p=0.014], with no effect on gene expression of inflammatory markers or angiotensin receptors (p>0.3). Ang‐(1‐7) did not alter body mass [Ang‐(1‐7): 46.5±2.0 g, saline: 45.9±1.5 g, p=0.792] or body composition (p>0.2). Ang‐(1‐7) also improved insulin sensitivity [Ang‐(1‐7): −2896±593 AUC, saline:‐1171 ± 624 AUC, p=0.033] but did not affect fasting glucose levels [Ang‐(1‐7): 180±5 mg/dL, saline: 169±8 mg/dL, p=0.225] or glucose tolerance (p=0.202).ConclusionsChronic Ang‐(1‐7) treatment decreases blood pressure, cardiac hypertrophy, and markers of cardiac sympathetic activity as well as increases insulin sensitivity in aged mice. These beneficial cardiometabolic effects of Ang‐(1‐7) occurred in the absence of significant changes in body weight or composition. Overall, these data suggest that Ang‐(1‐7) may provide a novel pharmacological target to improve age‐related cardiovascular and metabolic risk.Support or Funding InformationNIH Pathway to Independence Award: R00 HL122507 to A.C. ArnoldAHA Postdoctoral Fellowship: 18POST33960087 to A.J. Miller
Progesterone induced blocking factor improves blood pressure, mitochondrial dysfunction and reactive oxygen species in response to sFlt‐1 induced hypertension during pregnancy
Authors: Deer Evangeline , Jones Jalisa , Comely Kyliegh , Cornelius Denise C. , Ibrahim Tarek , Vaka Venkata Ramana , Franks Michael , LaMarca Babbette , Amaral Lorena M. 
Published: April 2020

Preeclampsia (PE) is characterized by new onset hypertension in association with placental ischemia, reduced fetal weight, elevated soluble fms‐like tyrosine kinase‐1 (sFlt‐1) and placental mitochondrial (mt) dysfunction and oxidative stress (ROS). Infusion of sFlt‐1 causes hypertension and endothelial and renal dysfunction in pregnant rodent models of PE. However a role for sFlt‐1 in causing mt dysfunction and ROS is unknown. Progesterone induced blocking factor (PIBF), is a product of progesterone signaling in early pregnant which controls pro‐inflammatory processes and thus markers of endothelial dysfunction. We have previously shown that progesterone supplementation improves markers of endothelial dysfunction in the RUPP rat. Moreover, we have more recently shown that PIBF lowers blood pressure in the RUPP rat model of PE. This study was designed to test two hypothesis. We first wanted to determine a role for sFlt‐1 to cause mt dysfunction and ROS during pregnancy. We next hypothesized the PIBF supplementation would lower in sFlt‐1 induced hypertension during pregnancy and improve mt function in response to sFlt‐1 during pregnancy. sFlt‐1 was infused into normal pregnant (NP) Sprague‐Dawley rats (3.7 μg·kg−1·day−1 for 6 days, gestation days 13–19) in the presence or absence of PIBF (2.0 μg/mL) administered intraperitoneal on GD 15 to normal pregnant (NP) rats. Mean arterial blood pressure (MAP) and placental mt ROS were measured on GD 19. Infusion of sFlt‐1 into NP rats increased MAP 112 ±2 (n=11) compared with control NP rats 98±2 mmHg (n=15, p<0.05). Administration of PIBF reduced MAP to 100±1 mmHg in the presence of sFlt‐1 (n=5, p<0.05). Mt ROS in placenta was 108±6 in NP (n=4), 429±32 in NP+ sFlt‐1 (n=3, p<0.05), and significantly reduced to 234±15 in NP+ sFlt‐1+ PIBF (n=3, p<0.05). State 3 respiration, which is indicative of ATP production, was reduced in placentas of sFlt‐1 infused rats versus NP, but was improved with PIBF. Overall, our study implicates a role for sFlt‐1 to reduce placental mt function during pregnancy and that supplementation of PIBF improved placental mt function which was associated with improved blood pressures. Our data indicate the importance of improved progesterone signaling to restore placental function in the presence of sFlt‐1 which led to improved hypertension and therefore should be considered a potential therapeutic for PE.Support or Funding InformationNIH Grants: RO1HD067541‐06 and P20GM121334; T32HL105324