bluuebunny/crossref_metadata_2025_split huggingface dataset:
Environmental noise aggravates oxidative DNA damage, granulocyte oxidative burst and nitrate resistance in Ogg1−/− mice
Authors: Oelze Matthias Walter , Kvandova Miroslava , Filippou Konstantina , Steven Sebastian , Frenis Katie , Kalinovic Sanela , Vijacic-Mirski Ksenija , Stamm Paul , Hosseinabadi Majid Bagheri , Dovinova Ima , Epe Bernd , Münzel Thomas , Kröller-Schön Swenja , Daiber Andreas 
Published: April 2020

BackgroundLarge epidemiological studies point towards a link between the incidence of arterial hypertension, ischemic heart disease, metabolic disease and exposure to traffic noise, supporting its role as an independent cardiovascular risk factor. We characterized the underlying molecular mechanisms leading to noise‐dependent adverse effects on the vasculature in an animal model of aircraft noise exposure identifying oxidative stress and inflammation as central players in mediating vascular dysfunction. Here, we want to study the impact of noise‐induced oxidative DNA damage on vascular function in DNA‐repair deficient Ogg1−/− (8‐oxoguanine glycosylase knockout) mice.Methods and ResultsOgg1−/− and corresponding wild type mice were exposed to aircraft noise (peak sound levels of 85 dB(A) and mean sound level of 72 dB(A)) for 4 days. We could show that 4 days noise exposure induced significant degree of endothelial dysfunction in wild type and Ogg1−/− mice. In contrast, endothelium‐independent relaxation was only impaired in noise‐exposed Ogg1−/−, indicating the presence of nitrate resistance in noise‐exposed Ogg1−/− mice. The cardioprotective, aldehyde detoxifying enzyme mitochondrial aldehyde dehydrogenase (ALDH‐2), which is necessary for nitroglycerin bioactivation, was upregulated only in the noise‐exposed Ogg1−/− mice, which may indicate the compensatory upregulation of dysfunctional ALDH‐2. Aircraft noise exposure caused oxidative DNA damage (8‐oxo‐deoxyguanosin) and elevated NOX‐2 expression in wild type mice with synergistic increases in Ogg1−/− mice (shown by immunohistochemistry). A similar pattern was found for oxidative burst of blood leukocytes and other markers of oxidative stress (4‐hydroxynonenal, 3‐nitrotyrosine) and inflammation (cyclooxygenase‐2).ConclusionThe finding that chronic noise exposure causes oxidative DNA damage in mice is worrisome since these potential mutagenic lesions could contribute to cancer progression. Human field studies have to demonstrate whether oxidative DNA damage is also found in urban populations with high levels of noise exposure as recently shown for workers with high occupational noise exposure.Support or Funding InformationWe gratefully acknowledge the financial support by the Naturwissenschaftlich‐Medizinisches Forschungszentrum (NMFZ) of the Johannes Gutenberg University Mainz (to B.E. and A.D.), the Boehringer Ingelheim Foundation „Novel and neglected cardiovascular risk factors: molecular mechanisms and therapeutic implications” is gratefully acknowledged (to S.S., T.M. and A.D.) and a postdoctoral research stipend for a scientific stay in Mainz by the Slovak Academy of Science (to M.K.).
Anti‐Oxidant Treatment Improves Aortic Vascular Compliance In Fructose‐Induced Salt‐Sensitive Hypertension in Male Rats
Authors: Levanovich Peter Eric , Komnenov Dragana , Perecki Natalia , Wu Min , Rossi Noreen F. 
Published: April 2020

High fructose diet increases renal reactive oxygen species (ROS) production which enhances renal sodium reabsorption and renin secretion, thereby increasing extracellular volume and vasoconstriction. With concurrent high sodium consumption, hypertension occurs prior to the development of frank metabolic syndrome. Prolonged hypertension causes vascular dysfunction by reducing compliance, leading to end‐organ damage to the heart and kidneys. Daily treatment with Tempol®, a ROS scavenger, prevents increases in mean arterial pressure (MAP) associated with a fructose and high salt (FHS) diet, indicating a pathologic role of ROS in this model. We hypothesized that daily administration of Tempol® ameliorates the reduction in vascular compliance and abnormalities in cardiac function associated with FHS induced hypertension. Male Sprague‐Dawley rats were fed either 20% glucose or 20% fructose in drinking water with normal (0.4%) salt chow for one week followed by 3 weeks of normal or high (4.0%) salt chow. Half of each group was supplemented with Tempol®, 15 mg/400 g body weight daily in drinking water, beginning at one week after fructose or glucose feeding. Hemodynamics were monitored by telemetry. Ultrasonography was performed under isoflurane anesthesia. The change in MAP from baseline in the FHS group was significantly elevated compared with controls (4.98 vs. 0.16 mmHg, respectively; P < 0.01), and Tempol® administration prevented this increase. Despite increased MAP, plasma renin activity in FHS rats was not suppressed. Ultrasonography revealed significantly decreased aortic compliance, measured by pulse wave velocity (P < 0.01), and significantly increased renal resistive index (RRI) in the FHS group compared to glucose controls (P < 0.05). Tempol® administration reversed the increase in pulse wave velocity but had no effect on RRI. Systolic left ventricular function by echocardiographic assessment did not change but diastolic function was impaired. This was evidenced by decreased ratio of early to late transmitral filling velocity (P < 0.01). Tempol® did not restore mitral flow patterns to control levels. These findings indicate that the decrease in vascular compliance associated with FHS diet in male rats is, at least in part, mediated by oxidative stress. Other mechanisms seem to be at play in mediating the abnormal renovascular changes.Support or Funding InformationVA Merit Award (RX000851) awarded to Noreen F. Rossi, MD. Peter Levanovich is supported in part by the Detroit Cardiovascular Training Program (NIH T32: HL 120822)
Determining conformational dynamics of the allosteric site in HRas R97G
Authors: Ojeda Monica , Johnson Christian , Mattos Carla 
Published: April 2020

HRas is a small GTPase that acts as a molecular switch for the Raf/MEK/ERK pathway that leads to cell proliferation, survival, and differentiation. Oncogenic mutations in HRas renders it unable to hydrolyze GTP to GDP, resulting in constitutive activation of HRas and cell proliferation. Additionally, HRas contains an allosteric site that binds Ca2+ and a negatively charged ligand, promoting helix 3 to shift towards helix 4 and the ordering of switch II. This conformation is associated with intrinsic hydrolysis of HRas in the presence of Raf. Experiments have been performed in our lab to study the effects of point mutations in the allosteric site of HRas on intrinsic hydrolysis constants. When compared to wild‐type, HRas R97G demonstrated increased intrinsic hydrolysis when alone and a significant decrease when in the presence of Raf. Molecular dynamics simulations were utilized to further gain insight on how the dynamics of the allosteric site differed in HRas R97G compared to the wild‐type. These simulations demonstrated increased flexibility in the upper region of helix 3, with greater accessibility to the catalytic conformation for GTP hydrolysis.Support or Funding InformationNSF MCB 1244203
C1orf27 controls the ER‐to‐Golgi traffic of GPCRs via direct interaction
Authors: Xu Xin , Qiu Lifen , Jiang Chunyi , Wu Guangyu 
Published: April 2020

C1orf27 is a human homolog of odorant response abnormal 4 which was originally identified as an endoplasmic reticulum (ER) membrane protein to regulate the delivery of odorant G‐protein‐coupled receptors (GPCRs) to cilia in C. elegans chemosensory neurons. However, its functions in the trafficking of mammalian GPCRs remain poorly understood. Here, we used α2‐adrenergic receptors (ARs) as models to define the role of C1orf27 in the biosynthesis and anterograde trafficking of GPCRs and elucidate the underlying mechanisms in several mammalian cells. We found that siRNA‐mediated depletion of C1orf27 significantly reduced the cell surface expression of α2A‐ and α2B‐ARs and in parallel receptor‐mediated signaling as measured by MAPK activation. Confocal microscopy revealed that the receptors were restricted in the perinuclear region and extensively co‐localized with the ER marker calregulin in C1orf27 siRNA‐treated cells. In contrast, C1orf27 knockdown did not affect the ER‐Golgi‐cell surface transport of non‐GPCR plasma membrane proteins, such as vesicular stomatitis virus glycoprotein. We also found that C1orf27 directly interacted with both receptors and the interactions were ionic in nature. Interestingly, two C1orf27‐binding sites were identified in the third intracellular loop (ICL3) and the C‐terminus of α2A‐AR, whereas only one binding domain was found in the ICL3 of α2B‐AR. Furthermore, the deletion of C1orf27‐binding domains clearly impaired the cell surface transport of the receptors, suggesting an important role of the interaction in receptor forward trafficking. Altogether these data have demonstrated that C1orf27 selectively modulates the ER‐to‐Golgi traffic of GPCRs which is likely mediated through direct interaction.Support or Funding InformationNIH R01 GM119815
An Ultrasensitive Fluorescent Biosensor for Multi‐modal Kinase Activity Detection and High‐resolution Imaging in Awake Mice
Authors: Mehta Sohum , Zhang Jin-Fan , Liu Bian , Hong Ingie , Tenner Brian , Lin Wei , Huganir Richard L. , Zhang Jin 
Published: April 2020

Fluorescent protein (FP)‐based kinase activity biosensors are powerful tools for probing the spatiotemporal dynamics of signaling pathways in living cells. Kinase activity imaging is frequently used to unravel the molecular regulation of Protein Kinase A (PKA), a prototypical kinase that integrates multiple pathways to orchestrate diverse physiological processes. Yet the limited sensitivity of most kinase biosensors restricts their reliable application in physiologically relevant in vivo systems or clinically useful high‐content detection modalities. We previously developed an excitation‐ratiometric PKA activity reporter (ExRai‐AKAR) which utilizes a PKA substrate peptide and phosphoamino acid‐binding forkhead‐associated 1 (FHA1) domain to modulate the peak excitation wavelength of cpEGFP in response to phosphorylation by PKA. Here, we report the further development of a new, ultrasensitive excitation‐ratiometric PKA activity reporter (ExRai‐AKAR2) obtained via high‐throughput linker library screening. This powerful and versatile molecular tool enables sensitive and rapid monitoring of live‐cell PKA activity across multiple fluorescence detection modalities, including plate reading, cell sorting, and one‐ or two‐photon imaging. In vivo visual cortex imaging in awake mice reveals highly dynamic neuronal PKA activity rapidly recruited by forced locomotion. The performance of ExRai‐AKAR2 promises to successfully expand kinase activity imaging beyond the petri dish.Support or Funding InformationThis work was supported by Brain Initiative Grant R01 MH111516 and by R35 CA197622, R01 DK073368, and R01 GM111665 (to J.Z.).
Role of transporters and S100A8/A9 in paclitaxel‐induced peripheral neuropathy
Authors: Li Yang , Zavorka Thomas Megan E. , Leblanc Alix F. , Huang Kevin M. , Chen Mingqing , Sprowl Jason A. , Gibson Alice A. , Lustburg Maryam M. , Sparreboom Alex , Hu Shuiying 
Published: April 2020

BACKGROUNDPaclitaxel (PTX) is a widely used anticancer drug that causes a debilitating, dose‐limiting acute and chronic peripheral neuropathy (PIPN) in up to 80% of patients. Although the mechanistic basis of this side effect remains largely unknown, we previously identified transport by OATP1B‐type carriers as a contributor to acute and chronic forms of peripheral neuropathy associated with PTX, and this effect can be ameliorated by the OATP1B2 inhibitor, nilotinib, without compromising anti‐tumor efficacy. Prior investigations have also shown that PTX induces the toll‐like receptor 4 (TLR4) in neurons and activation of TLR4 is associated with PIPN. In this study, we will explore potential effect of drug metabolism or transporter mediated drug‐drug interactions (DDIs) on PIPN, and novel mechanism involving S100A8/A9 proteins, as important mediators of inflammatory response in PIPN.METHODSIn vivo toxicity studies were performed in wild‐type, OATP1B2‐deficient, P‐gp‐deficient, CYP3A4‐deficient and S100A9‐deficient mice receiving PTX. Standard assessments of mechanical‐induced allodynia with Von Frey Hairs were conducted to evaluate PIPN at timed intervals before, during, and after treatment. Relationships between OATP1B2 and S100A9 gene or protein expression with or without PTX treatment was tested in mice with a deficiency of either OATP1B2 or S100A9.RESULTSSince PTX is substrate of P‐gp and metabolized by CYP3A4, the influence of P‐pg or CYP3A4 on PIPN was tested in mouse models deficient of either CYP3A4 or P‐gp. Our results suggest that neither CYP3A4 nor P‐gp deficient mice showed differences in sensitivity to mechanical stimuli compared to wild‐type mice after an i.v. dose of PTX. Interestingly, while OATP1B2 and S100A9 gene or protein expression are independently regulated at baseline, expression of S100A9 was dramatically upregulated in the dorsal root ganglia (DRG) of wild‐type mice after administration of a single dose of PTX. Since PIPN is functionally dependent on OATP1B2, which is sensitive to inhibition by small molecule inhibitors such as nilotinib without negatively affecting the anti‐tumor efficacy of PTX, we next evaluated the direct role of S100A9 in acute PIPN, and found that S100A9 deficiency protects against paclitaxel‐induced mechanical allodynia.CONCLUSIONOur preliminary results suggest that OATP1B2 inhibitor nilotinib could protect PIPN in a P‐gp and CYP3A4 independent way. Genetic knockout of S100A9 protects against acute forms of PIPN. Further studies are warranted to determine similar protective effects can be mediated by S100A8 deficiency, and to evaluate the translational utility of S100A8/A9 inhibitors as modulators of PIPN.
Many hands make light work, or too many cooks spoil the soup? Lessons from the development of the BASIL biochemistry laboratory curriculum through a diverse multi‐institutional collaboration.
Authors: Pikaart Michael 
Published: April 2020

The BASIL (Biochemistry Authentic Scientific Investigation Laboratory) curriculum is driven by a consortium of biochemistry and computational instructors. BASIL is a protein biochemistry Course‐based Undergraduate Research Experience (CURE) in which students discover enzymatic function of previously uncharacterized proteins. It combines computational and wet‐lab protein science to deliver foundational concepts within the undergraduate biochemistry teaching lab. BASIL came together in 2015 as instructors sought to integrate genuine research into the biochemistry laboratory course to broaden the impact of teaching through research.In order to maximize impact, BASIL has been intentionally developed as a collaborative effort across institutions diverse in type, size, and location nationwide. This presentation summarizes lessons learned from building this collaboration, including the following aspects.

(1) Establishing the group: Participants were identified and recruited for the most part at national meetings such as Experimental Biology/ASBMB and others.
(2) Funding: Development of the BASIL curriculum was supported by NSF IUSE as a collaborative research proposal.
(3) Communication: Group members interact through videoconferencing on a weekly basis, and at national/regional meetings.
(4) Teaching resources: The BASIL curriculum has been published as a free and open‐source resource, supplemented by online teaching suggestions.
(5) Assessment: The effectiveness of this curriculum has been assessed through student surveys shared across campuses and surveys of faculty to monitor success of (and obstacles to) implementation at each home campus.
(7) Dissemination and recruitment: Availability of the BASIL resources has been published, and additional faculty/campuses continue to join as BASIL implementors.These are presented with the goal of sharing successful (and not‐so‐successful) strategies for collaborating across campuses as a way to stimulate effective student learning while promoting faculty development and well‐being through this team model.Support or Funding InformationNSF IUSE 1709278
Regulation of the ER stress response by a mitochondrial microprotein
Authors: Chu Qian , Martinez Thomas F. , Novak Sammy Weiser , Donaldson Cynthia J. , Tan Dan , Vaughan Joan M. , Chang Tina , Diedrich Jolene K. , Andrade Leo , Kim Andrew , Zhang Tong , Manor Uri , Saghatelian Alan 
Published: April 2020

Cellular homeostasis relies on a dedicated and coordinated response to a variety of stresses. In the endoplasmic reticulum (ER) the accumulation of unfolded proteins triggers the unfolded protein response (UPR), a conserved stress response aimed at mitigating damage and returning cells to homeostasis. Dysregulation of UPR underlies many debilitating diseases. Here, we discover that a previously uncharacterized 54‐amino acid microprotein PIGBOS regulates UPR. PIGBOS localizes to the mitochondrial outer membrane where it interacts with the ER protein CLCC1 at ER–mitochondria contact sites. Functional studies reveal that the loss of PIGBOS leads to heightened UPR and increased cell death. The characterization of PIGBOS reveals an undiscovered role for a mitochondrial protein, in this case a microprotein, in the regulation of UPR originating in the ER. This study demonstrates microproteins to be an unappreciated class of genes that are critical for inter‐organelle communication, homeostasis, and cell survival.Support or Funding InformationThis research was supported by NIH/NIGMS (R01GM102491, A.S.), the Leona M. and Harry B. Helmsley Charitable Trust grant (A.S.), Dr. Frederick Paulsen Chair/Ferring Pharmaceuticals (A.S.), the George E. Hewitt Foundation for medical research (Q.C.), the Anderson Foundation (Q.C.), NIH F32 fellowship (GM123685, T.F.M.) and the Pioneer Fellowship(D.T.). Imaging work was supported by the Waitt Advanced Biophotonics Core Facility of the Salk Institute with funding from NIH‐NCI CCSG: P30 014195, NINDS Neuroscience Core Grant: NS072031 and the Waitt Foundation. Proteomics studies were supported by the Mass Spectrometry Core of the Salk Institute with funding from the NIH with an NCI Cancer Center Support Grant P30 (CA014195).
After exertional heat stroke, is cooling to <40°C sufficient? Revisiting an established protocol in a mouse model
Authors: Alzahrani Jamal Mohammed , Garcia Christian K. , Murray Kevin O. , Robinson Gerard P. , Sheikh Laila A. , Laitano Orlando , Clanton Thomas L. 
Published: April 2020

Exertional Heat Stroke (EHS) is a life‐threatening illness that can lead to multisystem tissue damage and increased mortality. To attenuate these consequences, immediate cooling to <40°C is the current recommendation. It is believed that the rate of cooling is a key factor in preventing tissue injury; however, much is unknown. In this study, we have used a model which allows us to test the effect of different cooling rates on recovery from EHS.METHODSThirteen female mice were randomly assigned to one of two groups, a slow‐cooling group (SC) and the rapid‐cooling group (RC). Both groups underwent emitter implantation surgery which allowed us to monitor core temperature (Tc) in real time. After two weeks of recovery, mice were introduced to free running wheels inside their cages for another two weeks. After that, mice received forced running wheel training for one week. On the day of EHS exposure, mice were placed inside a chamber, kept at 37.5°C/40% humidity. Mice ran at closely controlled speeds until they reached symptom limitation (i.e. loss of consciousness at ~42.3°C). The RC group was immediately taken outside the chamber and allowed to recover at room temperature ~23°C. However, the SC group was kept inside the running wheel enclosure until consciousness (~9 min) and then allowed to recover for another 2 hours inside their home cage at 37.5°C.RESULTSThe total thermal area averaged higher in the SC group, but did not reach statistical significance (P=0.1291). The time mice spent >39.5°C to the maximum Tc (Tc,max), and from Tc,max to 39.5°C was not statistically different. However, the SC group exhibited greater mortality than the RC (P=0.012). In the SC group, the time spent to cool from Tc,max to 37.5°C was statistically significant (P<0.0025). Also, the time from Tc,max to the minimum core temperature was statistically significant (P<0.0001).CONCLUSIONSIn conclusion, this model helped us to distinguish the effect of different cooling rates on mortality. The data suggests that quickly cooling to only 40°C may be insufficient to avoid heat injury or death, but cooling to 37.5°C may improve outcomes.Support or Funding InformationSupported by the BK and Betty Stevens Endowment and a Fellowship from King Saud University, Saudi Arabia
Stimulator of Interferon Genes (STING) Regulates Endothelial Healing and Reendothelialization Following Angioplasty‐Mediated Vascular Injury
Authors: Anastasiou Marina , Carrillo-Salinas Francisco Javier , Smolgovsky Sasha , Aronovitz Mark , Kaur Kuljeet , Alcaide Pilar 
Published: April 2020

Angioplasty and stent implantation are common clinical interventions to improve blood flow in Peripheral Artery Disease, which is rising in prevalence in America. However, these interventions result in vascular endothelial damage, leading to the activation of signaling pathways that can have a major effect on short‐ and long‐term vessel health. We hypothesized that endothelial cell‐intrinsic Stimulator of Interferon Genes (STING), a crucial regulator of immune responses triggered by pathogen or self‐derived DNA, regulates reendothelialization and healing in response to vascular injury. To study whether STING‐mediated reendothelialization was an endothelial cell specific effect, in vitro wound healing assay was performed on WT and STING deficient primary mouse heart endothelial cells (MHEC) using live imaging and quantification with Cytation and IncuCyte. We found that, while WT MHEC completely covered the scratch area within 24 hours of initial scratch, only 75.59% ± 5.8 of the initial wound was healed in STING deficient MHEC monolayers. Furthermore, our in vivo experiments using the wire injury‐mediated experimental model of angioplasty in mice followed by Evans Blue Dye staining to distinguish the areas with denuded endothelium, demonstrate a decrease in reendothelialization in injured carotid arteries in STING deficient mice as compared to WT mice five days post injury (43.57% ± 9.79 area reendothelialized vs 95.16% ± 2.77, respectively). Seven days post wire injury, WT carotid arteries were completely healed, in contrast to those of STING deficient mice that remained unhealed (67.74% ± 5.80 of reendothelialized vessel area). Additionally, multi‐parameter flow cytometry analysis of the enzymatically digested arteries from the seven days post wire injury group demonstrated an increase in myeloid cell infiltration in WT mice, which was not observed in either the injured STING deficient arteries or in the uninjured contralateral WT carotid arteries, indicating an early localized and differential immune response likely affecting long term vascular remodeling. Our results support that endothelial STING contributes to endothelial healing in vitro and in vivo, and that myeloid cell infiltration in the carotid arteries is required for reendothelialization and may be responsible for lack of healing in STING deficient mice. Ongoing experiments aim to definitively quantify temporal recruitment of immune cells over the course of reendothelialization and chronic vascular remodeling following wire injury and the mechanisms involved in short‐ and long‐term vascular health.Support or Funding InformationThis work was supported by the NIH‐RO1‐HL123658