bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sex Differences in Urinary Angiotensinogen (uAGT) Excretion, Renal Function, and Systolic Blood Pressure in 2‐Kidney, 1‐Clip Hypertensive Rats
Authors: Bell Annie L. , Shao Weijian , Katsurada Akemi , Sato Ryosuke , Navar L. Gabriel 
Published: April 2020

Intrarenal renin‐angiotensin system (RAS) activation and tissue injury in nonclipped kidneys of 2‐kidney, 1‐clip (2K1C) hypertensive rats have been characterized in previous studies of male but not female rats. To evaluate possible sex differences in response to renovascular hypertension, urinary angiotensinogen (uAGT) excretion, arterial blood pressure, urinary protein excretion, and renal function were assessed in 2K1C female rats.Female (n=8) and male (n=6) Sprague Dawley rats underwent placement of a 0.2 mm silver clip on the left renal artery to simulate unilateral renal artery stenosis. BP was measured by tail‐cuff plethysmography, and 24‐hour urine volume and water intake were monitored via metabolic cages over 21 days. After this period, renal clearance studies were conducted in anesthetized rats, and urine was collected from each kidney separately over 30 minute intervals. Urine protein concentration was determined by pyrogallol red method, and uAGT was measured by ELISA as an index of intrarenal RAS activity. Inulin and PAH concentrations were measured by spectrophotometry for calculation of glomerular filtration rate (GFR), renal plasma flow (RPF), and renal blood flow (RBF).21 days after renal artery clipping, systolic BP had risen to 176±8 mmHg from a baseline pressure of 120±1 mmHg, and urinary protein excretion increased to 20±5 mg/day in females. uAGT excretion was significantly increased from the baseline of 13 ng/day to 74 ng/day (507±127% increase). The nonclipped kidney showed 90±29% higher uAGT excretion compared to the clipped kidney, consistent with our previous findings in males. GFR and RPF were higher in nonclipped kidneys than in clipped kidneys but were greater than in corresponding measurements in males. GFR, RPF, and RBF were significantly higher in control rats compared to both clipped and nonclipped kidneys. Male rats showed significantly higher increases in uAGT, BP, and protein excretion.The present study demonstrates that BP, uAGT, and urinary protein excretion substantially increase after renal artery clipping in females, but the magnitude of the changes is markedly lower than in males. Additionally, the nonclipped kidney showed higher uAGT excretion compared to the clipped kidney. Although GFR, RPF, and RBF were significantly lower than control values for both clipped and nonclipped kidneys, these values were higher for females than males. These results demonstrated that female rats may be protected against AngII‐mediated renal injury and hypertension.Support or Funding InformationSupported by the ASPiRE Program at Tulane Univ Sch Medicine, New Orleans, LA and the Warren R. Bourgeois III, MD and Usha Ramadhyani Bourgeois, MD Student Research Endowed Fund at Tulane Univ Sch Medicine, New Orleans, LA
Commensal microbiota are essential for vascular contractility mediated by actin polymerization
Authors: Furtado Janara , Bearss Nicole R. , Edwards Jonnelle M. , Mell Blair , McCarthy Cameron G. , Vijay-Kumar Matam , Joe Bina , Wenceslau Camilla F. 
Published: April 2020

IntroductionCommensal microbiota within a holobiont contribute to the overall health of the host via mutualistic symbiosis. Disturbances in such symbiosis are prominently correlated with a variety of diseases affecting humans, including cardiovascular diseases, which are the number one contributors to mortality. Given that a hallmark of all cardiovascular diseases is changes in vascular function, we hypothesized that depleting microbiota from a holobiont would induce vascular dysfunction.MethodsTo test this hypothesis we used male inbred Sprague Dawley (SD) rats that were either germ‐free (GF) or re‐conventionalized germ‐free (GFC) rat (7 weeks‐old). Re‐conventionalization of GF rats were performed by co‐housing GF rats with conventionally‐raised SD rats for 10 days (1:1 ratio); this allows GF rats to receive a natural, continuous exposure to gut microbiota via coprophagy. Mesenteric resistance arteries were used to evaluate vascular function using a wire myograph. Vascular smooth muscle cells (VSMC) were cultured from aorta to evaluate cell migration via scratch assay; proliferation via IncuCyte; and cofilin phosphorylation (inactive form) via Western blot. We quantified cardiac hypertrophy by dividing the left ventricle weight by 26.73 + tibia length3. (Statistics: t‐test: p<0.05: *vs. GF, n=5–6/group)ResultsGF rats had a lower LV mass compared to GFC (GF: 6.9±0.2; GFC: 7.5±0.2*). Intriguingly, GF rats presented a reduction in both KCl‐and phenylephrine‐induced contraction and GFC ameliorated these responses [KCl: Emax (mN): GF: 4.8±1.0 vs. GFC: 8.5±1.1* and phenylephrine: Emax (mN): GF: 6.9±1.3 vs. GFC: 11.7±0.7*]. Given that no differences were observed in acetylcholine‐induced relaxation, we hypothesized that the hyporesponsiveness of the GF arteries were due to cytoskeleton dynamics. Indeed, we observed that GF rats present a reduction in cofilin phosphorylation (phosphoCofilin/TotalCofilin; AU: GF: 0.3±0.03 vs. GFC: 0.6±0.11*). Decreased phospho‐cofilin leads to its active form and subsequently actin depolymerization in VSMCs.ConclusionThis study shows that commensal bacteria are essential for normal structure and function of the vascular system of the host.Support or Funding InformationNIH: R00GM118885
Tissue distribution studies of the peptide‐bilayer coated magnetic nanobeads and delivery of an adducted peptide, to retard melanoma growth, in vivo
Authors: Natarajan Pavithra , Martinez Baltazar , Lingo Joshua D. , Fleming Sherry D. , Tomich John M. 
Published: April 2020

Self‐assembling peptides based delivery systems are gaining increased importance as they are highly biocompatible, biodegradable and tunable i.e. structure and function can be modified. Branched amphiphilic peptides (BAPs) self‐assemble to form bilayer delimited vesicles, called the Branched Amphiphilic Peptide Capsules (BAPCs), whose molecular architecture is similar to liposomes. BAPCs have been used to date to deliver encapsulated radionuclides and dsRNA and DNA electrostatically bound to the exterior of the capsules. Recently, we have synthesized magnetic nanoparticles with the BAP bilayer coating for studying their interaction with a biological system and for exploring applications in delivery. The magnetic nanoparticles are being used as quantification tools to study some aspects of the peptide bilayer interactions such as their route of uptake by cells in culture and to study their tissue distribution in mice, employing several routes of delivery. Recent in vitro studies using widely used techniques such as confocal imaging and flow cytometry alongside the quantification method using the newly developed BAP‐MNBs, demonstrate that they are readily taken up by cells in culture via more than one endocytic pathway. This study suggests that quantification using BAP‐MNBs is a reliable tool for studying BAPCs delivery system and associated molecules. Current studies involve studying the tissue distribution of BAP‐MNBs when injected intravenously in mice. The BAP‐MNBs are alongside being tested as delivery systems to deliver a therapeutic retro‐inverso D‐peptide (RD‐p9) known to retard the growth of melanoma tumors in mice. Our current studies in mice will provide insight into the fate of BAPCs in vivo, aiding future studies which aim to tailor and use BAPCs as well as BAP‐MNBs for specific applications.Support or Funding InformationPhoreus Biotechology Inc., Olathe, Kansas
Characterizing the role of POLQ variants in genomic instability and cancer
Authors: Ray Sreerupa , Sheld Francesca , Moen Avery , Avalos Lisbeth , Victorino Jorge , Towle-Weicksel Jamie 
Published: April 2020

Melanoma is the most serious form of skin cancer that arises from melanocytes. It is developed due to ultra‐violet (UV) radiation inducing DNA damage when left unrepaired lead to mutations, which lead to genomic instability. Unrepaired DNA, or its faulty repair, can lead to mutagenesis, which can propagate further through rounds of replication, eventually resulting in uncontrolled cell growth leading to cancer. However, cells have various kinds of DNA repair pathways, which includes specialized enzymes that can repair DNA. One of the most important DNA repair enzyme is DNA polymerase theta (Pol θ or POLQ). POLQ has been shown to be involved in double strand break repair through a non‐canonical DNA end‐joining pathway known as the micro‐homology mediated end‐joining (MMEJ). POLQ is a low fidelity DNA polymerase and it introduces various kinds of mutations during DNA repair. Previous reports show that breast cancer patients that overexpress POLQ tend to have poorer survival rate indicating a potential role for POLQ in cancer. In collaboration with the Yale SPORE in Skin Cancer and Rhode Island College, we have identified several variants of POLQ in melanoma patients. Here we demonstrate that the variants of POLQ induce genomic instability and have accumulations of DNA double strand breaks (DSBs). We show that POLQ variants are sensitive to DNA damaging agents like etoposide. In addition, biochemical studies indicate that some of these variants display a slower polymerization rate compared to wild‐type (WT) suggesting a different mechanism for nucleotide incorporation. Together, this suggests that the cancer‐associated variants lead to genomic instability. Our studies suggest that variants of POLQ alter the DNA repair landscape and has potential to impact cancer treatment.Support or Funding InformationThis research was supported by Linfield College Start‐Up Funds and Linfield College Faculty Student Research Collaborative Grant (FSRCG) to SR.
The Effects of Low Volume Versus High Volume Sled‐Push Training on Muscular Adaptation
Authors: Bernard Jeffrey , Liao Yi-Hung , Madrigal Christian , Levesque Joshua , Lee Sukho , Fraze Matthew , Del Toro Isaac 
Published: April 2020

PurposeTo compared the effects of low and high volume sled‐push resistance training on muscle strength, power, and body composition.MethodsTwenty‐four college students were recruited and matched based on baseline one‐repetition max (1‐RM) into 1 of 3 groups: 1) low volume (LV), 2) high volume (HV) resistance training groups, or 3) control (CON) (n = 8 per group). The LV training consisted of 5‐single repetitions pushing a weighted sled for 10 yards. The HV training consisted of a multi‐set protocol of 3‐sets of 5‐pushing a weighted sled for 10 yards. Training consisted of 3‐weekly workouts performed on nonconsecutive days for 6‐weeks. This study utilized a pre‐ and post‐test study design consisting of 1‐RM, Wingate power test, standing long jump, vertical jump, and body composition. The CON group only performed the pre‐ and post‐tests.ResultsFollowing 6‐weeks of training there was a similar, but significant, increase in 1‐RM for both training groups (pre‐test; LV= 500 ± 32.6 lbs vs HV 480 ± 42.9 lbs; post‐test; LV= 658.1 ± 33 lbs vs HV= 632.5 ± 35.3 lbs, P < .001). However, there were no improvements in the Wingate power test, standing long jump, vertical jump, or body composition between pre‐ and post‐tests for both training groups ( P > .05). There were no improvements for CON (P > .05).ConclusionThis study suggests that low volume resistance training is as effective as a high volume protocol for improving muscle strength. However, its effectiveness on muscle power and body composition are unclear.
Exploring Drug Resistance in HIV‐1 Protease using 3D Printed Models
Authors: Villeda Adan , Chivington Austin , Cobos Maximilian , Gastineau Thomas , Mares Cesar , Que Javier , Porter Lon A. , Novak Walter R.P. 
Published: April 2020

HIV roughly affects 38 million people around the world. The enzyme HIV‐1 protease is essential for replication and maturation of the virus. HIV‐1 protease activates a variety of important proteins including reverse transcriptase and RNase H. Darunavir, an HIV‐1 protease inhibitor, is the most potent antiretroviral drug to combat HIV to date. Despite the potency of Darunavir, HIV‐1 protease can develop mutations both inside and outside of its active site to render Darunavir and other inhibitors ineffective. Here we develop 3D printed models to illustrate the structural basis of drug resistance. Specifically, we model the mutations V32I and L76V in HIV‐1 protease. Hands‐on models that illustrate resistance mutations near the active site of HIV‐1 protease provide students with additional visual references to better understand drug resistance mechanisms and the design of novel inhibitors to effectively overcome these mutations.Support or Funding InformationThis work was funded in part by NSF‐IUSE 1725940 for the CREST Project.
Seeking Satiety: Dopamine Receptors 1,2 mRNA Expression in Channel Catfish
Authors: Wertz Blaine , Kostner Danica , White Taylor , Moyers Macy , Kobayashi Yass 
Published: April 2020

The dopaminergic nervous system regulates pathways of reward, pleasure, and satiety, and is therefore strongly related to food intake and subsequent eating behavior in mammals. Pathologies of this system have been well‐studied in both humans and rodents; however, little is known about this system in fish species, including channel catfish. To increase our understanding in the involvement of the dopaminergic nervous system in regulation of food intake in fish species, the expression of dopamine receptors 1 and 2 (DR1‐2) mRNA, and their modulation in response to changes in food intake was examined. Samples of the brain, hypothalamus, liver, muscle, spleen, kidney, and two segments of intestine were collected from three fish to examine the tissue distribution of DR1 and DR2 mRNA. In addition, the expression of DR1 and DR2 mRNA was examined in four segments of the channel catfish brain. DR1 and DR2 mRNA was detected across all tissues examined, with readily detectable expression in the brain and hypothalamus. Among brain segments, DR2 mRNA was most readily detectable in the hypothalamus and forebrain. Hypothalamic expression of DR1 and DR2 mRNA was unaffected by food exposure when samples were taken at one‐hour prior to feeding, at the time of feeding, and one‐hour post feeding. Currently, we are examining the expression of DR1 and DR2 mRNA in relation to feeding frequency, as well as prolonged fasting to further elucidate the relationship between food intake and dopaminergic neuron function.Support or Funding InformationK‐INBRE P20GM103418
BAT1 Alters Migration, Invasion and Pro‐Inflammatory Cytokines in Prostate Cancer
Authors: Garcia Aileen Marie , Viquez Olga , Roque Yarelis , Sanchez Maria , Martinez Magaly 
Published: April 2020

Prostate cancer (PCa) is the most frequently diagnosed non‐cutaneous cancer and the second cause of cancer related deaths in men. Though PCa is often treated with radical prostatectomy and radiation therapy, 15%–30% of patients may undergo biochemical recurrence. The current biomarkers are inadequate to identify recurrence, thus there is a demand for the development of biomarkers that can identify recurrence. In preliminary data, HLA‐B associated transcript 1 (BAT1) was down‐regulated in patients with PCa recurrence when compared with non‐recurrent PCa patients. BAT1 has been identified as an anti‐inflammatory gene in other types of diseases. The role of BAT1 in PCa has not been identified. We hypothesize that BAT1 expression will affect cell migration, invasion, and inflammatory cytokine expression in vitro and in vivo. Our objective was to identify the role of BAT1 expression in PCa. We down‐regulated and upregulated BAT1 in androgen independent PC3 and androgen dependent 22RV1 prostate cancer cell lines using BAT1 small‐interfering RNA (siRNA) and complementary DNA (cDNA) lentiviral particle infections, respectively. In vitro assays were performed to measure BAT1 protein expression, migration, invasion and gene expression in prostate cancer cells. Western blot analysis showed that BAT1 protein expression was significantly down‐regulated in PC3 and 22RV1 siRNA transfected cells and overexpressed in PC3 and 22RV1 cDNA infected cells when compared to control. BAT1 down‐regulation significantly increased cell migration and invasion when compared to control. In contrast, BAT1 overexpression significantly decreased cell migration and invasion when compared to control. Real time PCR (RT‐PCR) showed changes in expression of pro‐inflammatory cytokines (TNF‐α and IL‐6) and genes that regulate cell adhesion and migration (MMP10, MMP13 and TIMPs) in BAT1 overexpressed cells when compared to BAT1 siRNA cells. In vivo studies were performed using severe combined immunodeficient (SCID) orthotopic mice injected with 22RV1 cells previously transfected with BAT1 short hairpin ribonucleic acid (shRNA) or BAT1 cDNA. The cells were injected directly into the prostate and allowed to form tumors for analysis. Immunohistochemistry analysis showed increases in the expression of TNF‐α, IL‐6 and MMP10 in tumors developed from BAT1 shRNA treated mice when compared to tumors developed from BAT1 cDNA treated mice. These results suggest that BAT1 expression alters inflammation through the modulation of TNF‐α and IL‐6 expression and can eventually promote migration and invasion through the expression of MMPs and TIMPs.Support or Funding InformationThis work is supported by the NIGMS‐RISE Program #R25GM061838
The Structural and Functional Role of Vertebrate LARP6 N‐Terminal Region
Authors: Norbury Brianna , Lewis Karen 
Published: April 2020

La‐related protein 6 (LARP6) belongs to a superfamily of evolutionarily related RNA‐binding proteins. While other LARP families have been characterized more thoroughly, LARP6 is still poorly understood. The majority of the work to date for LARP6 focuses on its RNA binding activity, and has primarily studied the RNA binding domain, the “La Module”, in isolation. However, these studies may not reflect how the full‐length protein behaves in vivo. Previous work in our lab using the Danio rerio (zebrafish) LARP6 protein as a model system demonstrated that a domain adjacent to the La Module, called the “N‐terminal region” (NTR), may play an important role in the full‐length protein. To study the effect of this domain on the RNA binding activity, we created a new protein construct that contained only the NTR and the La Module while omitting the C‐terminal domain, dubbed the “ΔCTD.” The NTR has also been created with an attached His10‐SUMO tag, with the additional creation of the isolated tag to serve as a negative control for biochemical analyses. Several features have been compared between the La Module and our ΔCTD construct, with supporting evidence from the NTR. First, size exclusion chromatography and analytical ultracentrifugation demonstrated that, unlike the La Module, the ΔCTD forms dimers under nonreducing conditions. These dimers were resolved by the addition of reducing agent, suggesting that the dimerization was caused by a disulfide bond. RNA binding activity was measured using electrophoretic mobility shift assays and showed that, unexpectedly, the ΔCTD exhibited distinct binding behavior as compared to the isolated La Module. The ΔCTD appears to exhibit increased affinity to homopolymers poly‐A, poly‐U, and the endogenous stem‐loop ligand HsCOL1A1, but no binding was observed to poly‐C, showing that the NTR not only modulates the RNA binding activity, but also its ligand specificity. Circular dichroism of the ΔCTD and SUMO‐tagged NTR strongly indicates that the NTR is a random coil. Small‐angle X‐ray scattering (SAXS) analysis supports that the NTR is able to interact with the La Module to modulate RNA binding behavior.Support or Funding InformationNational Institutes of Health – National Institute of General Medical Sciences (R15GM119096), National Institute of Health project ALS‐ENABLE (P30 GM124169) and a High‐End Instrumentation Grant (S10OD018483)
Do Action Potentials in Vasopressin Magnocellular Neurons Back‐propagate from Soma to Dendrites to Evoke Neuropeptide Dendritic Release?
Authors: Zhang Meng , Stern Javier E. 
Published: April 2020

In addition to releasing their peptidergic cargo into the systemic circulation from their axons in the posterior pituitary, magnocellular neurosecretory vasopressin (VP) and oxytocin (OT) neurons in the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei also release these neuropeptides from their dendrites. Dendritic release of VP and OT play important functional roles, including recruitment and coordination of neurohumoral responses to an osmotic stimulation. Dendritic release of neuropeptides involves Ca2+‐dependent exocytosis, which in other neuronal types, including substantia nigra dopaminergic neurons, has been shown to be triggered by backpropagating action potentials (bAPs). However, it is at present unknown whether action potentials in VP neurons can back‐propagate from soma to dendrites, and whether they participate in dendritic release of VP. To directly test this, we performed simultaneous dual patch‐clamp electrophysiological recordings from somata (whole‐cell, current‐clamp) and dendrites (loose patch, voltage clamp) in identified VP neurons of eGFP‐VP rats. Various degrees and patters of somatic firing activity were evoked by direct current injection through the somatic patch pipette. Surprisingly, we found evidence for bAPs only in a very small proportion of recorded VP neurons (~20%, 91/435 recordings). When present, bAPs appeared as fast inward deflections that occurred within less than 1 millisecond from the evoked somatic AP. bAPs displayed high fidelity, even at relatively high somatic firing frequencies (up to 10 Hz), with almost no propagation failures observed within the measured distance from the soma. The presence or absence of bAPs was not dependent on neuronal input resistance, dendritic access resistance or dendritic distance from the soma (15–100 μM) Similarly, neither the number, frequency nor pattern (continuous or bursting) of somatic action potential had an influence on the incidence or properties of bAPs (p> 0.5 in all cases). Blockade of K+ channels with 4AP (5 mM n=11) or TEA (30 mM, n=7) did not enabled somatic action potentials to back‐propagate to the patched dendrite. Ongoing morphometric analysis of patched neurons aim at determining whether back‐propagation of action potential correlates with specific morphological features of these neurons. Taken together, our results indicate that despite being highly electrotonically compact, the vast majority of neurosecretory VP neurons do not show evidence for bAPS. These findings raise important questions regarding mechanisms underlying activity‐dependent dendritic release of neuropeptides in hypothalamic neurons.Support or Funding InformationNIH R01NS094640 to JES