bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ser14‐Rpn6/PSMD11 Phosphorylation Mediates the Activation of 26S Proteasomes by cAMP and Protects against Cardiac Proteotoxic Stress in Mice
Authors: Yang Liuqing , Parajuli Nirmal , Wu Penglong , Liu Jinbao , Wang Xuejun 
Published: May 2022

BackgroundMeans to enhance proteasome (PSM) functioning are highly sought. In cultured cells, cAMP‐dependent protein kinase (PKA) was shown to activate 26S PSMs by phosphorylating Ser14 of Rpn6/PSMD11 (a key PSM subunit), but this remains to be established in animals. Hence, we sought to test the hypothesis that Ser14‐Rpn6 phosphorylation (pS14‐Rpn6) mediates the activation of PSMs by PKA and reduces proteotoxicity in vivo (Figure 1).Methods and ResultsWe generated via CRSPR/Cas9 gene editing two mouse models, in which the codon for Ser14 in the Rpn6 gene was mutated to encode either Alanine (S14A) for blocking pS14‐Rpn6 or Aspartic acid (S14D) for mimicking this phosphorylation. Treatment with adenylate cyclase activator forskolin (5mg/kg, i.p.) increased cardiac pS14‐Rpn6 (2.52±0.08 vs. 1.0, p<0.001) and PSM activities (292.35±45.75 vs. 100, p<0.005) in wild type mice, but these were abolished by co‐treatment of PKA inhibitor H89 (15mg/kg). Forskolin did not show these effects in S14A mice, demonstrating that pS14‐Rpn6 is required for PKA to activate the PSM. Cross‐breeding transgenic GFPdgn (a proven substrate of the ubiquitin‐proteasome system [UPS]) into S14D mice revealed that S14D decreased cardiac GFPdgn proteins (0.81±0.08 vs. 1.0±0.08, p<0.05) without altering GFPdgn mRNA levels, indicating that S14D suffices to enhance UPS performance. Basal myocardial PSM activities were significantly higher in S14D mice compared to WT mice (1.52±0.38 vs. 1.0±0.16, p<0.005). To determine the effect of increasing pS14‐Rpn6 on proteotoxicity, we cross‐bred S14D into the CryAB‐R120G mice (R120G), a classic model of cardiac proteotoxicity. We detected that myocardial total (0.76±0.07 vs. 1.0±0.19, p<0.005) and insoluble (0.80±0.12 vs. 1.0±0.14, p<0.01) CryAB‐R120G protein levels were lower in S14D::R120G compared to WT::R120G mice (n=4 males+4 females/group), suggesting that PSM activation by PKA decreases the accumulation and aggregation of misfolded proteins in vivo. Echocardiography further revealed that S14D significantly attenuated the decrease of left ventricular (LV) ejection fraction (51.80±11.65 vs. 40.20±13.88 %, p<0.01), stroke volume (38.44±7.30 vs. 28.65±5.69 μl, p<0.005) and cardiac output (17.45±3.09 vs. 12.77±2.46 ml/min, p<0.005) of R120G mice at 6 months of age, which indicates that genetic mimicry of pS14‐Rpn6 ameliorates cardiac proteotoxicity.ConclusionsWe have established for the first time in vivo that pS14‐Rpn6 is required for activation of 26S PSMs by PKA and provides the first genetic evidence that PKA‐induced PSM activation protects against proteotoxicity in animals.
Mitochondrial oxidative phosphorylation fuels endothelial control of blood vessel tone
Authors: Wilson Calum , Lee Matthew D. , Buckley Charlotte , Zhang Xun , McCarron John G. 
Published: May 2022

Arteries and veins are lined by non‐proliferating endothelial cells that play a critical role in regulating blood flow. Endothelial cells also regulate tissue perfusion, metabolite exchange, and thrombosis. It is thought that endothelial cells rely on ATP generated via glycolysis to fuel each of these energy‐demanding processes. However, endothelial metabolism has mainly been studied in the context of proliferative cells in angiogenesis, and little is known about energy production in endothelial cells within the fully‐formed vascular wall.Using intact arteries isolated from rats and mice, we show that inhibiting mitochondrial oxidative phosphorylation at mitochondrial complex V disrupts calcium‐dependent, nitric oxide‐mediated endothelial cell control of vascular tone. Basal, mechanically‐activated, and agonist‐evoked calcium activity in intact artery endothelial cells are each prevented by inhibiting mitochondrial ATP synthesis. This effect is mimicked by blocking the transport of pyruvate, the master fuel for mitochondrial energy production, through the mitochondrial pyruvate carrier. The role for endothelial cell energy production is independent of species, sex, or vascular bed.These data show that mitochondrial ATP is necessary for the obligatory role of endothelial cells in the control of blood vessel diameter, and validate the idea of targeting endothelial cell metabolism to treat endothelial cell dysfunction in cardiovascular disease.
Neuroprotective Role of Hyperoxia During the Critical Period of Retinal Development in Normal Mice and Mice with Retinitis Pigmentosa
Authors: Villegas Monica Anne Faye , Gdovin Eric , Tilvawala Megha , Brahmbhatt Rushil , Sutera Christopher , Lecker Paul , Berstein Jake , Tsai E‐Jine , Martin Mark , Kim Jessica , Morrison Nathan , Gorski Grzegorz , Antonello Zeus , George‐Weinstein Mindy , Bravo Nuevo Arturo 
Published: May 2022

BackgroundThe retina utilizes more oxygen than any other tissue in the body. Photoreceptors located in the retina's outer nuclear layer (ONL) are culled during the “critical period” of eye development when the number of photoreceptors exceeds the supply of oxygen. Retinitis Pigmentosa (RP) is a disease that first affects the rod photoreceptors, causing night blindness and reduced peripheral vision. In mice, RP often starts during the critical period. The objective of this study was to assess the effects of hyperoxia during the critical period on visual function and morphology of the retina in normal mice and mice with RP.MethodThirty five male and female wild‐type (C57BL/6J) mice and 15 mice with an rd1 mutation (C3H/HeJ), which causes a rapidly progressing form of retinitis pigmentosa, were exposed to hyperoxia (75% O₂), hypoxia (12% O₂), and normoxia (21% O₂) from postnatal (P) days 7 to 20. At P26, electroretinography (ERG) was used to assess the visual function of the photoreceptors (A‐wave) and the inner retinal cells (B‐wave). At P27, optical coherence tomography (OCT) and histology were used to observe retinal structure and the effects of the different O₂ groups. For histology, eyes were extracted at P10, P12, P16, and P28. Cell death was evaluated using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL).ResultsIn ERGs from C57 mice, both A‐wave and B‐wave amplitudes were significantly lower in the hypoxic condition indicating that hypoxia was detrimental to visual function compared to the normoxic group. OCT data showed that the hyperoxic groups of C57 mice had thicker ONLs compared to the other treatment groups. TUNEL cell counts in the ONL and INL indicated a significant decrease in apoptotic cells during the critical period in the hyperoxic group compared to the hypoxic group. In C3H mice, hyperoxia improved the A and B waves compared to untreated mice.DiscussionThis study suggests that administration of oxygen during the critical period of retinal development reduces the pruning of cells in the retina and increases visual function in both normal mice and mice with an aggressive form of RP. Oxygen treatment during the critical period of retinal development, which occurs from P8 to P16, may slow retinal degeneration in RP. The effects of hyperoxia on the completion of vascular development must be taken into account when considering the use of hyperoxia therapeutically.
Signaling Pathways Associated with miR‐106b mediated Radioresistance in the Treatment of Colorectal Cancer
Authors: Rubio‐Guevara Susana , Olascuaga‐Castillo Karyn , Cáceres‐Andonaire Elena , Altamirano‐Sarmiento Dan , Caballero‐Aquiño Olga , Mantilla‐Rodríguez Elena , Hilario‐Vargas Julio , Berezovski Maxim , Morgado‐Díaz José Andrés 
Published: May 2022

Tumor radioresistance, or the lack of control of certain tumors with this treatment, can result in locoregional recurrence; therefore, there is great interest in understanding the underlying biology and developing strategies to overcome this problem. MicroRNAs (miRNAs, miRs) are small non‐coding RNAs that regulate gene expression at the post‐transcriptional level and participate in cancer invasion, progression, metastasis, and therapeutic resistance. Emerging evidence indicates that miRNAs play a critical role in modulating key cellular pathways that mediate response to radiation, influencing radiosensitivity of cancer cells through interaction with other biological processes such as cell cycle checkpoints, apoptosis, autophagy, epithelialmesenchymal transition, and cancer stem cells. Today, most studies on patient data report different, results on the miRNAs evaluated for each tumor type, highlighting miR‐106b whose overexpression can determine radioresistance both in vitro and in vivo by inhibiting apoptosis and promotion of cell proliferation. The objective of this study was to find the signaling pathways involved with miR‐106b‐mediated radioresistance.MethodsCRC gene expression data sets were collected from the public database, The Cancer Genome Atlas (TCGA). In addition, web‐based tools were used to explore TCGA data, specifically that developed by the Memorial Sloan Kettering Cancer Center (MSK) cBioPortal for Cancer Genomics. The public site cBioPortal is hosted at the MSK Molecular Oncology Center, in which the term “colorectal cancer” was searched and 12 studies were selected, creating a single combined study which has 4341 patients and 4488 samples, within which the search for miR‐106b was carried out and the signaling pathways involved with the expression mediated by this miRNA were obtained.ResultsThe following signaling pathways involved with miR‐106b were obtained: WNT, TP53, TGF‐Beta, RTK‐RAS, PI3K, NRF2, NOTCH, MYC, HIPPO, and its influence on the cell cycle was also noted.ConclusionmiRNAs have been shown to play an important role in the regulation of CRC radio resistance, by controlling various signaling pathways, including cell cycle, proliferation, apoptosis, and DNA damage repair. Furthermore, these results are consistent with recent data that have shown that selective modulation of miRNA activity can improve the response to radiotherapy, providing an innovative antitumor approach based on miRNA‐related gene therapy. Therefore, miRNAs could also serve as targets for the development of new therapeutic strategies to overcome radiation resistance in CRC.
Pedagogical Lessons Learned During the Time of COVID‐19
Authors: Ortiz Phillip , Dehner Carolyn 
Published: May 2022

As COVID‐19 swept the globe colleges and universities opted to suspend face‐to‐face lectures and laboratories as a step to slow the spread of this disease. Rather than cancel their courses instructors worked to deliver content and assess learning via distance learning and online pedagogies. Since this shift occurred mid‐semester and unexpectedly, instructors and curriculum designers needed to make the transition quickly and smoothly. The urgency of the situation was not a call for lowering standards or changing learning outcomes. Rather, it was an opportunity to revisit our courses with a focus on the critical learning outcomes.During their course transitions many instructors discovered that creating coursework for an online setting is not a transcriptional process, in which only the medium by which the content delivery differed, but a translational one in which they needed to use an entirely different set of tools. For example, while creating video lecture captures or transcribing the text of the lectures may be the quickest way to make the transition, in the long‐term instructors may have used readings from a variety of sources; relatively brief written, audio or video “mini‐lectures”; different textbooks, open educational resources, and other content sources; and importantly, incorporated frequent assessments with rapid feedback.Delivering laboratory content via online modalities can be especially challenging. Moving it online may have been an opportunity to reconfigure an experience that often requires that students focus much of their efforts on data collection, to one that emphasizes hypothesis formulation, experimental design, and data analysis. It is possible that when this crisis passes some of the insights gained in moving courses online will be useful in enhancing learning in traditional settings.To help educators transition to online teaching, the leadership of Biochemistry and Molecular Biology Education (BAMBEd) compiled a freely accessible virtual issue that may serve as useful resources for BMB educators who are converting their face‐to‐face courses to the online modality. That content may be accessed at: http://IUBMB.onlinelibrary.Wiley.com/doi/toc/10.1002/(ISSN)1539‐3429.Teaching‐COVID‐19That virtual collection also marked BAMBEd's commitment to continue to expand this knowledge base and to make new resources available to the community of educators quickly. To that end, we welcomed brief rapid communications from educators on creative and innovative approaches. In addition, we also welcomed submissions by the leadership of professional organizations as they make statements of educational priorities and offer guidance to the field.
Assessing the Effectiveness of flowcharts as a teaching and learning tool
Authors: King Ethan , Zimmermann Anthony E. , Bose Diptiman 
Published: May 2022

The Integrated Cardiovascular Acute Care disease module in the second professional year at Western New England University, College of Pharmacy and Health Sciences, introduces disease states' pharmacotherapeutics such as coronary artery disease, shock, and arrhythmias. To improve learner understanding and application of this material, we introduced the use of flowcharts in the module. Flowcharts are skeletal outlines of key concepts that help summarize acute cardiovascular diseases by combining pharmacological concepts with evidence‐based pharmacotherapeutic decision‐making. This study aims to evaluate the effectiveness of flowcharts as an instructional tool in the therapeutics module.A skeletal outline of the flowcharts was provided to the students on the College’s learning management site. The instructors completed the charts using a doc‐cam system during class at the end of each disease state. A pre‐ and post‐survey was administered to evaluate the effectiveness of these flowcharts. The study was initiated in Spring 2019 and data was collected until Spring 2021.A pre‐survey showed that 83% (n=45) agreed that flowchart use would improve their knowledge; the post‐survey showed that 91% (n=60) agreed that flowcharts improved their overall knowledge in the course. While 90% (n=60) agreed that flowcharts improved their understanding of pharmacology, 95% of the surveyed participants agreed that flowcharts helped them understand the role of drugs in pharmacotherapy. Furthermore, 96% (n=60) of the participants agreed that they would like to see more use of flowcharts in the other pharmacotherapeutics modules. A topic‐wise comparison of the average scores demonstrated significant differences in the mean±SD score on summative assessments for the classes that received flowcharts versus those that did not use flowcharts.In order to gain insight into how the students use flowcharts for learning, we also conducted focus‐group interviews of the students using predesigned questions. A total of 16 students were interviewed in groups of 4‐5. Thematic coding of the interview transcripts is currently being performed. However, preliminary 1st and 2nd round coding identified emerging themes such as “visually useful,” “summarizes material,” “helps retain and memorize”, “helps in exam” etc.The results from our study indicate that the use of flowcharts improved content delivery and application in an acute care cardiovascular disease pharmacotherapeutics module. The flowcharts provide the students with a decision‐making framework to make informed therapeutic decisions. We believe that the use of these flowcharts benefit student learning and understanding and can be used in other disease state modules.
Upper airways dysmorphology and obstructive sleeping apnea in Down syndrome
Authors: Martínez Abadías Neus , Echeverry Luis Miguel , Maréchal Laura , Giménez Sandra M. , Sevillano Xavier M. , Fortea Juan M. , Heuzé Yann 
Published: May 2022

Down syndrome (DS) is a complex genetic disorder caused by trisomy of chromosome 21 that alters the development of multiple systems, including craniofacial dysmorphologies. Mandibular hypoplasia, macroglossia and midfacial flatness are associated with higher prevalence of malocclusion and mouth breathing in DS. In this study, we assessed the morphology of the upper airways and tested whether alterations in the size and shape of the upper airways were associated with Obstructive Sleep Apnea (OSA), which compromises the quality of life in individuals with DS.To quantify differences in volume and shape of the upper airways, we obtained 3D reconstructions from head magnetic resonances scans in a cross‐sectional sample of 246 adult individuals, including 159 control euploid individuals and 87 individuals diagnosed with DS. Polysomnography tests were available in a subsample of individuals with DS, and AHI Index scores representing the number of apnea and hypopnea events per hour of sleep were used to indicate the severity of sleep apnea.Geometric Morphometric analyses based on the 3D coordinates of 15 anatomical landmarks located on the surface of the upper airways showed significant shape differences between DS and control individuals (P‐value=0.001), involving a retraction of the whole upper airways structure and a change in the angular inclination of the oropharynx region in individuals with DS. Volumetric analyses confirmed a significant 40% reduction in total upper airway volume in DS (P‐value<0.0001). Multivariate regression analysis showed that AHI scores explained 10.1% of upper airways morphological variation in DS, with a significant positive correlation between the shape and volume of the upper airways and the severity of OSA. Our study suggests that morphological biomarkers assessed from non‐invasive magnetic resonance images can be useful in diagnosing OSA, a sleep disturbance with high prevalence in DS that if treated correctly, can improve the quality of life of people with DS and prevent the appearance of early dementia.
Smooth Muscle‐Specific Deletion of Neuropilin‐1 Increases Vascular Contractility and Blood Pressure
Authors: Balasubbramanian Dakshnapriya , Lambrinos George , Cristofaro Vivian , Bigger‐Allen Alexander , Wang Beibei , Gao Yao , Chen Hong , Adam Rosalyn M. , Sullivan Maryrose P. , Bielenberg Diane R. 
Published: May 2022

IntroductionNeuropilin‐1 (NRP1) is a transmembrane glycoprotein that acts as a co‐receptor for the Class 3 Semaphorin (SEMA) ligand SEMA3A and mediates the inhibition of RhoA signaling. NRP1 is expressed in vascular smooth muscle cells (VSMCs), however, its role in regulating VSMC contractility and blood pressure in vivo is unknown.HypothesisWe hypothesize that SEMA3A‐NRP1 signaling is essential in mitigating RhoA activity in VSMCs and that loss of NRP1 increases vascular tone and blood pressure in vivo.MethodsTo study the role of NRP1 in SMCs, mice with inducible, smooth muscle cell‐specific deletion of NRP1 (SM22a‐CreERT2; Nrp1flox/flox) were generated. Following recombination using 4‐hydroxy tamoxifen (SM‐NRP1 KO) in male and female adult mice (8‐12 weeks), systolic blood pressure (SBP) was measured using a tail cuff and compared to age‐ and sex‐matched mice that did not receive tamoxifen (control). Vascular reactivity to various contractile agonists was measured using ex vivo tension myography.ResultsSBP was significantly increased in SM‐NRP1 KO mice following recombination compared to control mice (SBP: 136.5 ± 10.9 vs 112.9 ± 5.6 mmHg; p=0.0006). Aortas of SM‐NRP1 KO mice displayed significantly enhanced contractile response to phenylephrine, KCl, and the thromboxane agonist U44619, and reduced relaxation to the endothelium‐independent vasodilator sodium nitroprusside. In vitro, treatment of murine primary aortic VSMC with SEMA3A decreased angiotensin II‐induced Rho‐GTP activation. Gene expression of Sema3a in the aorta was downregulated in two models of hypertension, angiotensin II‐dependent and genetic model of hypertension, as identified by qPCR and transcriptomic analysis respectively, further supporting the role of SEMA3A‐NRP1 signaling in basal blood pressure regulation. Additionally, control and SM‐NRP1 KO mice (starting at 2 weeks post‐recombination) were administered angiotensin II (490 ng/kg/day) for 4 weeks. While there was no significant difference in SBP at weeks 1 and 2, SM‐NRP1 KO mice had significantly lower SBP at weeks 3 and 4 following angiotensin II infusion compared to controls (Week 4 SBP: 150 ± 1.4 vs 130.5 ± 2.5 mmHg; p=0.02), suggesting a low ejection fraction and cardiac dysfunction in these mice. In support of this observation, gene expression of atrial natriuretic peptide was increased (p=0.06) in hearts of angiotensin II‐infused SM‐NRP1 KO mice. Preliminary analyses indicated an increase in aortic medial thickness at baseline and an increase in aortic adventitial collagen following angiotensin II infusion in SM‐NRP1 KO mice compared to controls.ConclusionTogether, our data point to the role of NRP1 as a novel regulator of basal vascular tone and blood pressure, and the loss of NRP1 leads to the onset of hypertension and exacerbates pathological vascular remodeling and cardiac dysfunction.
Investigating the Protective Antibody‐Circumsporozoite Complex of Plasmodium falciparum and its Role in Vaccine Development
Authors: Dobelhoff Katie , Dobelhoff Ben , Fraley James , Suder Karen C. , Replogle Jessica B. 
Published: May 2022

Malaria is a parasitic disease associated with high fevers, shaking chills, and flu‐like symptoms. It is typically spread by mosquitoes and caused by infection with Plasmodium falciparum (Pf). Due to its high transmission rate and metastatic properties, malaria is a major public health concern. Insecticide resistance has led to alternative methods of disease management, such as vaccine development. The circumsporozoite protein of Plasmodium falciparum malaria is the foundation for many malaria vaccines, which are designed to block the transmission of Pf. The most promising vaccine candidate is RTS,S, a recombinant protein designed around the central NANP repeat and C‐terminal region of circumsporozoite protein. The repeat region consists of 19 repeating NANP amino acid sequences, allowing multiple antibodies to bind simultaneously. Molecular dynamics simulations and crystallography structures show the importance of the NANP amino acid sequences for binding. The Summit Country Day School MSOE Center for Biomolecular Modeling MAPS Team used 3D modeling and printing technology to examine the structure‐function relationship of the human Fab688 circumsporozoite complex and its associated binding of NANP repeat peptides to protective antibodies. Antibodies that bind to the fully extended peptide are perpendicular to the heavy chain‐light chain interface and rely on the first NANP repeat that follows the only NPDP sequence in the junctional region of Pf circumsporozoite protein, as this arrangement favors free energy contributions and results in superior protection. Junctional peptide antibody protection may relate to the ability to cross‐react with the NANP repeat region. Amino acids Asn11 and Asn13 on the junctional peptide form a hydrogen bond network with Fab668. This web of hydrogen bonds, which connects the residues that form the binding pocket, stabilizes it and contributes to stronger binding.Fab688 binds to the NANP repeat region with equal or better affinity, shedding light on the affinity and specificity for the junctional region of circumsporozoite. This research lays the framework for future advancements in vaccines by revealing important targets and thereby limiting the spread of infectious diseases.