bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immediate and Late Effects of Anabolic Androgenic Steroids on Testicular Morphology of Rats
Authors: Felix‐Patrício Bruno , Guarçoni Thaissa Lange , Marchon Roger Gaspar , Medeiros Jorge Luiz , Gregório Bianca Martins , de Souza Diogo Benchimol 
Published: June 2021

Background and objectiveThe anabolic androgen steroids are indiscriminately used and in high doses by amateurs and professional athletes. Although its effects on different organs are well known, few is reported regarding the testicular morphological effects (both immediate and late) of these steroids. Thus, the objective of this study was to investigate the effects of supraphysiologic doses of nandrolone decanoate on the testicular morphology of rats; and investigate if these effects would be transitory or permanent.MethodsTwenty‐eight adult male Wistar rats were assigned into four groups. For eight weeks, control groups received placebo (oleaginous vehicle) injections while treated groups received nandrolone decanoate (10mg/Kg, intramuscularly, once a week). Groups control‐immediate (CI) and treated‐immediate (TI) were killed one week after the last injection. Groups control‐late (CL) and treated‐late (TL) were killed after 12 weeks from the last injection. Body weight, testicular weight and volume were measured. The testicle was processed for histomorphometrical analysis, and epithelium height and tubular area were obtained. Data was compared by Student's t test considering p<0.05 as significant.ResultsNo difference was observed regarding body weight. Also, testicular weight and volume was similar among groups TI and CI. However, group TL showed a reduction of 44% and 48% in testicular weight and volume, respectively, in comparison to CL. The tubular area was reduced in both TI (by 22%) and TL (by 46%), in comparison to their control groups. Further, epithelium height was reduced by 9% in group TI.ConclusionThe use of supraphysiologic doses of nandrolone decanoate promotes testicular morphological alterations in rats. These alterations are not reversible after discontinuing the treatment and may cause permanent subfertility.Support or Funding InformationFAPERJ, CNPq, CAPESThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Long‐term dietary nitrite supplementation is effective in reducing cardiovascular disease risk factors in older, obese rats
Authors: Kadel Jacob L. , Van Schaeffer G , Bates Melissa L. , Sindler Amy L. 
Published: June 2021

Cardiovascular diseases (CVD) persist as the leading cause of death. CVD risk increases with advancing age, and is further exacerbated in settings of obesity. Sodium nitrite supplementation improves cardiovascular abnormalities in several models of dysfunction. Given the association between aging, obesity and CVD risk, we tested the hypothesis that long‐term sodium nitrite supplementation would reduce CVD risk factors in older, obese rats. At 6 months of age, male Fisher 344 rats were randomly split into two groups; animals fed HFD (60%(Research Diets); n = 10) with or without nitrite‐supplemented water (50 mg/L; HFD‐NO2−; n = 10) for 10 months. Age‐matched rats fed regular rodent chow (2920X (Teklad); n = 10) served as controls (CON).As expected, rats on a HFD for 10 months were heavier vs. CON (598 ± 11 vs. 432 ±10 gms; p <0.0001), which nitrite attenuated (533 ± 14 gms; p = 0.0021). DEXA scans revealed that the reduced body weight in the HFD‐NO2− rats was accompanied with reduced body fat % (28 ± 0.6 vs. HFD 31 ± 0.4%; p = 0.0046), while having no effect on lean % (HFD‐NO2− : 48.1 ± 0.4 vs. HFD: 47.5 ± 0.2%; p = 0.30).Compared to CON, HFD rats had increased systolic blood pressure (SBP: 120 ± 3.81 vs. 136 ± 5.40 mmHg; p = 0.03), diastolic blood pressure (DBP: 75 ± 2.49 vs. 87 ± 4.24 mmHg; p = 0.04) and mean arterial pressure (MAP: 90 ± 2.71 vs. 103 ± 4.53 mmHg; p =0.03), which was completely prevented in HFD‐NO2− (SBP: 114 ± 4.07 mmHg; p = 0.004, DBP: 74 ± 4.15 mmHg; p = 0.04, and MAP: 87 ± 4.05 mmHg; p = 0.016, all vs. HFD). Furthermore, HFD‐NO2− rats had reduced fasting glucose when compared to HFD rats (97 vs. 1.3 vs. 104 ± 2.6 mg/dL; p = 0.02).Taken together, these data demonstrate that nitrite supplementation may have therapeutic potential for treating and/or preventing CVD risk factors that are associated with aging and obesity.Support or Funding InformationSupported by NIH K01AG047626, University of Iowa College of Liberal Arts and Sciences Old Gold Summer Fellowship, and University of Iowa Department of Health and Human PhysiologyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Menopausal Hormone Treatments on Intravascular Cellular Activation and Development of White Matter Hyperintensities in Healthy Postmenopausal Women
Authors: Jayachandran Muthuvel , Lahr Brian D. , Miller Virginia M. , Kantarci Kejal 
Published: June 2021

ObjectiveBlood thrombogenicity and vascular inflammation may influence development of white matter hyperintensities (WMH) in the brain. This study aimed to determine if menopausal hormone treatments (MHT) modified that association in recently menopausal women who were enrolled in the Kronos Early Estrogen Prevention Study (KEEPS) at Mayo Clinic.MethodsWomen (n=95) underwent MRI prior to and at 18, 36 and 48 months after randomization to either transdermal 17β‐estradiol (tE2, n=30), conjugated equine estrogen (oCEE, n=29) both with progesterone for 12 days per month or placebo pills and patch (PBO, n=36). Fasting blood was analyzed for a set of 14 activation markers of platelets and endothelial cells, inflammation, and thrombogenicity. Principal components (PC) analysis was used to reduce dimensionality of these markers, and the first 5 PC identified were then assessed for association with treatment and changes in WMH.ResultsWMH increased in all groups over the 48 months of treatment with greater increases in the oCEE compared to tE2 and PBO (P=0.015). The 5 PC explained 62% of the variation in cellular activation markers. A global test of these 5 PC revealed a borderline association with treatment group (P=0.059), with the PC reflecting total platelet count, platelet microaggregates and ATP secretion showing higher scores in the oCEE compared to the other groups (P=0.006). The change in WMH over the 48 months of MHT showed an overall association with the 5 PC (P=0.026), the most prominent of which was the first PC reflecting vascular inflammation by number of blood microveicles (MV) from activated monocytes and endothelial cells and MV positive for intercellular adhesion molecule 1 (P=0.001). There was no evidence that this association differed by MHT.ConclusionMarkers of blood thrombogenicity and inflammation may influence development of WMH in the brain but this relationship may be influenced by factors other than the type of MHT used in the KEEPS study.Support or Funding InformationNIH P50 AG44170This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Malaria derived extracellular vesicles inhibit neutrophils ROS production and NETs formation
Authors: Babatunde Kehinde Adebayo , Walch Michael , Fellay Isabel , Kharoubi‐Hess Solange , Filgueira Luis , Ghiran Ionita , Mantel Pierre‐Yves 
Published: June 2021

IntroductionA dysfunctional innate immune response is believed to provide immune evasion of the malaria parasites, but also to cause increased susceptibility to bacterial infections. Neutrophils are the most abundant cells found in the blood circulation in direct contact with parasite infected red extracellular vesicles (EVs) derived from iRBCs, containing both parasite and host materials, including microRNAs, modulate neutrophil response by transferring regulatory microRNAs.Methods and ResultsWe have previously reported that malaria EVs contain miR451a, a microRNA that is known to regulate neutrophil activity when EVs are phagocytized by neutrophils. Thus we monitored the uptake of EVs by neutrophils by fluorescence microscopy, confocal microscopy and real‐time polymerase chain reaction techniques as well as potential pathways involved in their uptakes by neutrophils. Furthermore, our data demonstrated the influence of malaria induced EVs on human neutrophil functions in vitro by inhibiting their ability to produce ROS and suppressing cytokine secretion. Neutrophils are known to produce neutrophil extracellular traps (NETs) during immune response to pathogens; therefore, we quantified the production of NETs with incubation of EVs.ConclusionOur data indicate that EVs are actively taken up by neutrophils to deliver miR­451, thereby interfering with their capacity to kill bacteria by inhibiting ROS and NETs formation. We describe a new mechanism of cellular communication between parasites and the host immune system. While EVs might increase tolerance to the parasites, they dramatically affect the resistance to a co­infection by bacteria. The elucidation of the immune regulatory role of EVs might lead to the development of new therapies.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metformin inhibits autophagy and mitophagy in cardiomyocytes
Authors: Chang Michael , Pinkhasova Polina , Kobayashi Tamayo , Patel Rahul , Cohen Max , Mehta Puja , Zhang Yawen , Kobayashi Satoru , Liang Qiangrong 
Published: June 2021

Metformin, a first‐line medication for the treatment of type 2 diabetes mellitus, has been suggested by several clinical studies to be cardioprotective. However, the mechanisms underlying metformin's cardioprotection remain debated. One current hypothesis proposes that metformin protects the heart by increasing autophagy in cardiomyocytes, leading to increased removal of protein aggregates and damaged organelles including mitochondria. In this study, we tested this hypothesis using both in vitro and in vivo models. Rat H9C2 cardiac myoblasts were treated with 3 mM metformin for 24 hours and autophagy/mitophagy were determined by Western blot analysis of LC3‐II, a well‐established marker of autophagic vesicles. Surprisingly, metformin significantly reduced LC3‐II levels in either total cell lysates or mitochondrial fractions; and lysosomal inhibitors increased LC3‐II in metformin‐treated cells to a less degree as compared with vehicle‐treated cells. These results suggest that metformin inhibited rather than stimulated autophagy and mitophagy, contrary to the current belief. This was confirmed by a novel dual fluorescent mitophagy reporter showing reduced mitochondrial fragments being degraded in the lysosomes in the presence of metformin. The reduced mitophagy was also found in the heart from the mitophagy reporter mice that received 200 mg/kg of metformin through gavage. Collectively, these results indicate that the cardioprotective benefit of metformin observed in many different models may be through mechanisms other than increased autophagy and mitophagy.Support or Funding InformationThis work was supported by the National Institutes of Health National Center for Research Resources.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assembly of Fragment Ends After PCR: An Efficient and Accurate Multi‐part DNA Assembly Method for Large DNA Sequence
Authors: Lin Yibin 
Published: June 2021

Recent development of DNA assembly technologies has spurred myriad advances in synthetic biology. But new tools are always required for complicated scenarios. Here, we have developed an alternative DNA assembly method named AFEAP cloning (Assembly of Fragment Ends After PCR), which allows scarless, modular, and reliable construction of biological pathways and circuits from basic genetic parts. The method requires two‐round of PCRs followed by ligation of the sticky ends of DNA fragments. The first PCR yields linear DNA fragments and is followed by a second asymmetric (one primer) PCR that inserts overlapping overhangs at both sides of each DNA fragment. The resulting products of the second PCR are then annealed and ligated with T4 DNA ligase, followed by bacterial transformation to yield the desired plasmids. We characterized its capability and limitations and demonstrated its application to assemble DNA with varying scenarios. Under the optimized conditions, AFEAP cloning allows assembly of an 8 kb plasmid from 1–12 fragments with high accuracy (between 80 and 100%), and 8.0, 11.6, 19.6, 28, and 35.6 kb plasmids from five fragments at 91.67, 91.67, 88.33, 86.33, and 81.67% fidelity, respectively. AFEAP cloning also is capable to construct bacterial artificial chromosome (BAC, 200 kb) with a fidelity of 46.7%. AFEAP cloning provides a powerful, efficient, seamless, and sequence‐independent DNA assembly tool for multiple fragments up to 12 and large DNA up to 200 kb that expands synthetic biologist's toolbox.Support or Funding InformationThis work was supported by National Modern Agriculture Industry Technique Systems (CARS‐02) to Jingao Dong and Starting Grant from Hebei Agricultural University to Fanli Zeng (grant number ZD201622).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Epigenetic Factors in Age‐Dependent Changes in Drosophila Melanogaster Visual Behavior and Photoreceptor Neuron Gene Expression.
Authors: Escobedo Spencer 
Published: June 2021

Age is a major risk factor for ocular disease. The goal of our lab is to understand the gene expression changes that occur with age. Our previous work shows that there is a visual senescence, the decrease in visual function with age, in Drosophila melanogaster. We have also shown that this age‐dependent decrease in visual function correlates with changes in gene expression. Epigenetic factors have been proposed to play a key role in maintaining ocular function with age. However, the epigenetic mechanisms that are involved in age‐dependent gene expression changes have yet to be identified. To address these questions we use Drosophila photoreceptor neurons as a model of the aging eye. We have conducted an RNAi screen of photoreceptor‐specific knockdown of these epigenetic factors which enables us to identify if they are necessary for visual senescence. Factors that are necessary for visual senescence can be further assessed for their role in gene regulation through photoreceptor‐specific RNA sequencing. This will identify the target genes of these factors, revealing potential mechanisms of age‐dependent regulation of gene expression. One specific factor is Brahma, the catalytic subunit of BAF, an ATP‐dependent chromatin remodeling complex. Preliminary data in the lab shows an enhanced visual senescence in Drosophila expressing a dominant negative Brahma in photoreceptors. Our current work is identifying the gene expression changes in these flies by RNA sequencing.Support or Funding InformationResearch reported in this poster was supported by the National Eye Institute of the National Institutes of Health under award number R01EY024905This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
JARID2 Regulates Skeletal Muscle Differentiation through Regulation of Canonical Wnt Signaling Pathway
Authors: Adhikari Abhinav , Davie Judy 
Published: June 2021

Myogenesis is a tightly regulated process that is dependent on the precise and dynamic integration of signals from various signaling pathways including the Notch, Sonic hedgehog and Wnt signaling pathways. The Wnt signaling pathway is an essential signaling pathway that has been shown to have diverse roles during both embryonic muscle development and adult muscle regeneration. In this study, we sought to understand the role of JARID2 ‐ an evolutionarily conserved, inactive histone demethylase that often has been shown to function in association with a transcription repressive complex, polycomb repressive complex‐2 (PRC2), during muscle differentiation. Utilizing the widely used C2C12 cell model system, we report that JARID2 is required for muscle differentiation and it regulates muscle differentiation through modulation of Wnt signaling. We found that JARID2 regulates the myogenic regulatory factors (MRFs), MyoD and myogenin. We discovered that JARID2/PRC2 not only regulates expression of the MRFs directly through tri‐methylation of lysine 27 of histone 3 (H3K27me3) but also indirectly, through regulation of canonical Wnt signaling. JARID2 controls MRF expression indirectly through direct repression of the Wnt antagonist gene, secreted frizzled‐related protein‐1 (Sfrp1). We found that activation of canonical Wnt signaling through inhibition of GSK3b rescues the differentiation block of JARID2 depleted cells through the restoration of MyoD and myogenin expression. JARID2 depleted cells showed a reduction in the nuclear levels of the canonical‐Wnt signaling pathway effector, b‐catenin, and we found that inhibition of GSK3b induced translocation of b‐catenin into the nucleus of these cells. Using chromatin immunoprecipitation (ChIP) assays, we also show that b‐catenin positively regulates the transcription of MyoD through direct binding in the distal regulatory region (DRR) of the MyoD promoter. Thus, we establish here that JARID2 and the PRC2 complex are essential regulators of the initiation of skeletal muscle differentiation through modulation of the Wnt signaling pathway. In myoblasts, PRC2 represses the MRFs but also represses Wnt antagonists, which allows the Wnt pathway to activate MyoD and initiate differentiation.Support or Funding InformationNIH/NIAMS AR068622This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Preventing Autoimmunity in Transcription‐Dependent CRISPR‐Cas Systems
Authors: Johnson Kaitlin , Learn Brian , Bailey Scott 
Published: June 2021

The CRISPR‐Cas systems are a collection of RNA‐based adaptive immune systems that protect many prokaryotes from invasion by viruses and plasmids. CRISPR‐Cas systems are very diverse and can be organized into six types and more than 15 subtypes. Type III systems are unique as their targeting mechanism requires transcription of the target DNA. Type III effector complexes use small CRISPR RNAs (crRNAs) to identify complementary transcripts. Transcript binding simultaneously activates two nuclease domains, leading to the degradation of both the bound transcript and adjacent DNA. Like all other immune systems, CRISPR‐Cas systems must differentiate between self and non‐self in order to prevent autoimmunity, which could arise from targeting of the host CRISPR‐array. Type III systems use two mechanisms to identify self‐transcripts. Studies of the Type III‐A system from Staphylococcus epidermidis revealed that base pairing between the crRNA and the sequence flanking the 3′ end of RNA target site prevents autoimmunity. However, a study of the Type III‐B system from Pyrococcus furiosus revealed that a short sequence motif flanking the RNA target sequence is required for immunity and the absence of this motif in CRISPR arrays prevents autoimmunity. Neither of these mechanisms is well understood biochemically and it is not clear if these mechanisms are conserved within the subtypes. To address these points, we monitored RNA‐activated DNA cleavage by the Type III‐B effector complex from Thermotoga maritima with purified components. A series of RNA transcripts containing various target 3′‐flanking sequences were tested for their ability to activate DNA cleavage. We find that two factors are important in preventing autoimmunity in the T. maritima Type III‐B system: (1) base pairing between the crRNA and the target 3′‐flanking sequence of the transcript and (2) the presence of a G, which does not base pair with the crRNA, immediately adjacent to the 3′‐end of the RNA target sequence. These data suggest that the mechanisms used to avoid autoimmunity by Type III systems are not subtype‐specific and that identification of self transcripts is complex, involving base pairing and specific sequence recognition.Support or Funding InformationThis work was supported by National Institutes of Health grant GM097330 to S.B.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
NAD+ Bound N‐terminal Domain of CARDS Toxin Increases IL‐1β Secretion
Authors: Woods Allison Taylor , Segovia Jesus , Galaleldeen Ahmad 
Published: June 2021

Mycoplasma pneumoniae (Mp) is an atypical bacterium that is linked to various respiratory diseases such as walking pneumonia and asthma. Upon infection, Mp produces a 591‐aa virulence factor known as Community Acquired Respiratory Distress Syndrome Toxin (CARDS TX). The crystal structure of this 591‐aa cytotoxin reveals a triangular molecule comprised of an N‐terminal ADP‐ribosylating domain and a C‐terminal tandem β‐trefoil domain that is responsible for vacuolation of the host's cells. Based on structural and sequence homology to other ADP ribosylating toxins, the NAD+ binding site has been predicted, yet none of the published structures of the CARDS toxin contain NAD+. The loop connecting the N− and the C‐termini is anchored using a disulfide bond that surrounds the NAD+ binding site which might sterically hinder NAD+ binding. An N‐terminal truncation mutant (NTTM) lacking the disulfide bond has been cloned and the protein was expressed, purified and crystallization trials are underway. The NTTM was also used to treat U937 cells in presence and absence of NAD+ to test the effect of the NTTM on cytokine secretion. The data show that NTTM causes an increase in the secretion of IL‐1β and TNF‐α in presence of NAD+ suggesting an increase in the inflammatory response to the NTTM in presence of NAD+.Support or Funding InformationSt. Mary's University Biaggini Funds and The San Antonio Livestock Expedition FundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.